Salivary Features of Periodontitis and Gingivitis in Type 2 Diabetes Mellitus

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Ebersole, Sreenatha S. Kirakodu, Xiaohua D. Zhang, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4391317/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 28 Dec, 2024 Read the published version in Scientific Reports → Version 1 posted 13 You are reading this latest preprint version Abstract Type 2 diabetes mellitus (T2DM) is associated with cellular abnormalities, tissue and organ dysfunctions, and periodontitis. This investigation examined the relationship between the oral microbiome and salivary biomarkers in T2DM patients with or without periodontitis. This cohort (35–80 years) included systemically healthy non-periodontitis (NP; n = 31), T2DM without periodontitis (DWoP; n = 32) and T2DM with periodontitis (DWP; n = 29). The oral microbiome [Operational Taxonomic Units (OTUs)] (16s rRNA sequencing) and targeted host salivary biomarkers (immunoassays) were assessed. We identified 47 OTUs that were significantly different in abundance between NP samples and any disease subset or between disease subgroups. The most unique microbiome patterns were observed in the DWP group. Differences in genera/species abundance were also observed when T2DM patients were stratified by extent of periodontal inflammation and disease (i.e., generalized versus localized gingivitis/periodontitis). Salivary biomarkers showed significant elevations in MMP-8, MMP-9, resistin, IL-1β, IL-6, IFNα, and BAFF (THFSR13b) comparing generalized to localized periodontitis. Salivary analytes showed significant positive correlations with specific microbiome members, predominantly in DWP patients. Odds ratio analyses reinforced that a panel of biologic markers (IL-6, MMP-8) and bacteria (e.g., Bacteroidetes, Fusobacteria, Spirochaetes ) discriminated the severity and extent of periodontal disease in this diabetic population. Biological sciences/Immunology Biological sciences/Microbiology Health sciences/Biomarkers Health sciences/Diseases diabetes microbiome inflammation periodontitis saliva Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 INTRODUCTION Type 2 diabetes mellitus (T2DM) is a chronic inflammatory disease with elevated systemic biomolecules of inflammation that are central to the pathogenesis of the disease. 1 – 5 Beyond the identification of periodontitis as a sequelae of T2DM, localized inflammatory proteins are routinely increased in the gingival crevicular fluid and saliva of periodontitis affected T2DM patients. 6 – 10 Within this oral environment the host responses are driven by, and reflect, an altered microbiome with dysbiotic changes of opportunistic pathogens and pathobionts revealing a lesional process. 11 – 15 As variations in the characteristics of the oral microbiome and associated host responses have been related to extrinsic and intrinsic (e.g., T2DM, age) effectors of the host ecosystem 16 – 19 , there remain gaps in our knowledge concerning the biologic mechanisms that are controlling these features in the oral cavity. Additionally, mechanical therapeutics in systemically normal individuals have been shown to substantially impact the microbiome and help transition its characteristics to one more consistent with periodontal health. 20 – 22 Nevertheless, existing data indicate that the prevalence and severity of periodontitis is increased in T2DM 23 – 27 . This investigation cross-tabulated the microbiome and biomarkers in saliva of T2DM patients with or without periodontitis. The findings provide insight into the unique features of localized and generalized gingivitis and periodontal disease in the presence of T2DM. RESULTS Oral clinical features in diabetes Figure 1 provides the periodontal characteristics and BMI relative to the various subgroups of T2DM and NP subjects. Only the NP group showed a significantly lower BMI compared to most of the T2DM groups (e.g., DWoP, DWP). In contrast and as expected, the clinical features of destructive disease in the generalized periodontitis patients were significantly different from both the gingivitis and localized periodontitis groups. Additionally, BOP showed significantly greater levels with increasing severity of disease. Of note, the majority of periodontally healthy individuals were in the NP group, and the majority of gingivitis cases were in the DWoP group. Based upon the disease features in affected patients, an additional perspective evaluated the characteristics of the microbiome and host response patterns related to the magnitude of disease. Microbiome characteristics in generalized and localized disease in diabetes Supplemental Fig. 1 provides a summary of the individual sample OTU reads for the NP, GG, LG, GP, and LP salivary microbiomes. Generally, similar numbers of overall reads were identified across the subgroups. In another report we identified the top 104 OTUs based upon levels across the three study groups (NP, DWoP, DWP), and showed that this portfolio of microorganisms provide 96–98% coverage of the microbiome reads in the samples 38 (Table 1 ). Figure 2 A presents the results for the individual NP subjects and shows Firmicutes (Bacillota) as the dominant phyla, with the relative abundance varying from ~ 40–80%. Bacteroidetes phyla was next most prevalent, being detected in 23 subjects. Proteobacteria was observed in the remaining six individuals. A subset of NP subjects demonstrated elevated levels of Spirochetes and four individuals had elevated Absconditabacteria in their salivary microbiome. Table 1 Operational taxonomic units (OTUs) included in the analysis. OTU Phyla Genera/Species Otu150 Absconditabacteria_(SR1) Absconditabacteria_(SR1)_[G-1] bacterium_HMT_345 Otu130 Absconditabacteria_(SR1) Absconditabacteria_(SR1)_[G-1] bacterium_HMT_875 Otu025 Actinobacteria Actinomyces graevenitzii Otu029 Actinobacteria Actinomyces sp._HMT_169 Otu007 Actinobacteria Actinomyces sp._HMT_180 Otu005 Actinobacteria Actinomyces_unclassified Otu049 Actinobacteria Atopobium_unclassified Otu106 Actinobacteria Bifidobacterium longum Otu053 Actinobacteria Corynebacterium matruchotii Otu014 Actinobacteria Rothia mucilaginosa Otu080 Actinobacteria Rothia_unclassified Otu105 Bacillota Selenomonas artemidis Otu163 Bacillota Selenomonas sp._HMT_126 Otu023 Bacillota Selenomonas sp._HMT_136 Otu034 Bacillota Selenomonas sputigena Otu019 Bacillota Selenomonas_unclassified Otu048 Bacillota Stomatobaculum longum Otu087 Bacillota Stomatobaculum sp._HMT_097 Otu064 Bacteroidetes Alloprevotella rava Otu058 Bacteroidetes Alloprevotella sp._HMT_308 Otu111 Bacteroidetes Alloprevotella sp._HMT_473 Otu042 Bacteroidetes Alloprevotella tannerae Otu147 Bacteroidetes Bacteroidales_[G-2] bacterium_HMT_274 Otu098 Bacteroidetes Bergeyella sp._HMT_322 Otu062 Bacteroidetes Capnocytophaga gingivalis Otu063 Bacteroidetes Capnocytophaga granulosa Otu065 Bacteroidetes Capnocytophaga leadbetteri Otu074 Bacteroidetes Capnocytophaga sputigena Otu083 Bacteroidetes Porphyromonas endodontalis Otu057 Bacteroidetes Porphyromonas gingivalis Otu038 Bacteroidetes Porphyromonas pasteri Otu173 Bacteroidetes Prevotella dentalis Otu054 Bacteroidetes Prevotella denticola Otu136 Bacteroidetes Prevotella intermedia Otu033 Bacteroidetes Prevotella nanceiensis Otu070 Bacteroidetes Prevotella nigrescens Otu018 Bacteroidetes Prevotella oris Otu068 Bacteroidetes Prevotella oulorum Otu017 Bacteroidetes Prevotella pallens Otu164 Bacteroidetes Prevotella pleuritidis Otu015 Bacteroidetes Prevotella salivae Otu092 Bacteroidetes Prevotella sp._HMT_300 Otu073 Bacteroidetes Prevotella sp._HMT_305 Otu011 Bacteroidetes Prevotella veroralis Otu001 Bacteroidetes Prevotella_unclassified Otu085 Bacteroidetes Tannerella sp._HMT_286 Otu067 Firmicutes Catonella morbi Otu135 Firmicutes Dialister invisus Otu031 Firmicutes Gemella morbillorum Otu027 Firmicutes Granulicatella adiacens Otu056 Firmicutes Lachnoanaerobaculum orale Otu125 Firmicutes Lachnoanaerobaculum umeaense Otu035 Firmicutes Lachnospiraceae_[G-2] bacterium_HMT_096 Otu022 Firmicutes Megasphaera micronuciformis Otu158 Firmicutes Mogibacterium vescum Otu252 Firmicutes Mycoplasma faucium Otu051 Firmicutes Oribacterium asaccharolyticum Otu052 Firmicutes Oribacterium_unclassified Otu112 Firmicutes Parvimonas micra Otu137 Firmicutes Peptostreptococcaceae_[XI][G-1]_unclassified Otu078 Firmicutes Peptostreptococcus stomatis Otu094 Firmicutes Ruminococcaceae_[G-1] bacterium_HMT_075 Otu129 Firmicutes Ruminococcaceae_[G-2] bacterium_HMT_085 Otu012 Firmicutes Streptococcus oralis_subsp._dentisani_clade_058 Otu008 Firmicutes Streptococcus parasanguinis_clade_411 Otu006 Firmicutes Streptococcus salivarius Otu004 Firmicutes Streptococcus_unclassified Otu003 Firmicutes Veillonella dispar Otu009 Firmicutes Veillonella parvula Otu119 Firmicutes Veillonella sp._HMT_780 Otu069 Firmicutes Veillonella sp._HMT_917 Otu002 Firmicutes Veillonella_unclassified Otu152 Firmicutes Veillonellaceae_[G-1] bacterium_HMT_155 Otu024 Fusobacteria Fusobacterium nucleatum_subsp._vincentii Otu059 Fusobacteria Fusobacterium sp._HMT_203 Otu013 Fusobacteria Fusobacterium_unclassified Otu044 Fusobacteria Leptotrichia hongkongensis Otu047 Fusobacteria Leptotrichia sp._HMT_212 Otu020 Fusobacteria Leptotrichia sp._HMT_215 Otu095 Fusobacteria Leptotrichia sp._HMT_218 Otu045 Fusobacteria Leptotrichia sp._HMT_221 Otu185 Fusobacteria Leptotrichia sp._HMT_223 Otu046 Fusobacteria Leptotrichia sp._HMT_392 Otu055 Fusobacteria Leptotrichia wadei Otu016 Fusobacteria Leptotrichia_unclassified Otu165 Fusobacteria Sneathia_unclassified Otu071 Proteobacteria Campylobacter rectus Otu171 Proteobacteria Campylobacter sp._HMT_044 Otu026 Proteobacteria Campylobacter_unclassified Otu091 Proteobacteria Escherichia coli Otu110 Proteobacteria Haemophilus influenzae Otu010 Proteobacteria Haemophilus_unclassified Otu115 Proteobacteria Kingella_unclassified Otu097 Proteobacteria Lautropia mirabilis Otu028 Proteobacteria Neisseria macacae Otu066 Proteobacteria Neisseria oralis Otu039 Proteobacteria Neisseria perflava Otu021 Proteobacteria Neisseria_unclassified Otu050 Proteobacteria Pasteurellaceae_unclassified Otu146 Spirochaetes Treponema denticola Otu099 Spirochaetes Treponema socranskii Otu156 Synergistetes Fretibacterium fastidiosum Otu123 Synergistetes Fretibacterium sp._HMT_361 Otu114 Synergistetes Fretibacterium_unclassified We next stratified the diabetic periodontitis (DWP) and gingivitis (DWoP) patients based on the clinical presentation (i.e., generalized or localized form) of periodontal inflammation and lesions to examine the phyla distribution. Firmicutes was the dominant phyla across the majority of DWoP patients with mean levels similar between generalized and localized gingivitis (Fig. 2 B). Three individuals showed Bacteroidetes as the dominant phyla. Generally, this phylum appeared more prevalent in the GG patients, while the Proteobacteria phylum was elevated across the LG subset. An elevated abundance was seen in LG for Absconditabacteria. Figure 2 C presents the phyla data for the DWP group with GP and LP. These generalized disease adults showed higher levels of Bacteroidetes, Fusobacteria, Proteobacteria, Spirochaetes , and Synergistetes compared to the localized disease group although the differences did not reach statistical significance. The evaluation of the microbiome differences was extended to focus on the individual microbial components in the various patient groups. Figure 3 illustrates the relative abundance of the targeted set of 104 OTUs in samples from the study population based on the level of oral disease (i.e., NP [healthy], LG, GG, LP, and GP). We identified 47 OTUs (Fig. 2 ) that were significantly different in abundance between NP and any disease subset or between disease subgroup categories (Table 2 ). There also were numerous OTU abundance differences (N = 18) between LG and LP. Seven bacterial species differences in abundance were observed between GG and GP, four bacterial species abundance differences were observed between LG and GG, and only one species ( P. gingivalis ) that was different in relative abundance between LP and GP. Table 2 Microbiome relative abundance differences between generalized and localized disease in T2DM patients. These taxa are significantly different between the groups determined by a t test (see Fig. 2 ). GG vs. LG GP vs. LP GG vs. GP LG vs. LP Prevotella _unclassified M. micronuciformis Prevotella sp._HMT_305 Bergeyella sp._HMT_322 P. gingivalis F. nucleatum_subsp._vincentii Prevotella nigrescens Rothia_unclassified Alloprevotella tannerae P. gingivalis Catonella morbi Fretibacterium fastidiosum Veillonella_unclassified Gemella morbillorum Prevotella nigrescens Stomatobaculum sp._HMT_097 Capnocytophaga gingivalis P. endodontalis Treponema socranskii Selenomonas sp._HMT_126 Prevotella intermedia Porphyromonas gingivalis Treponema denticola Fretibacterium fastidiosum Tannerella sp._HMT_286 Fretibacterium_unclassified Kingella_unclassified Bacteroidales_[G-2]_HMT_274 Prevotella sp._HMT_300 Mycoplasma faucium GG: generalized gingivitis, GP: generalized periodontitis, LG: localized gingivitis, LP: localized periodontitis. Salivary biomarkers, disease severity, and microbiome in diabetes Salivary concentrations of host response analytes by study group and periodontal disease category are depicted in Fig. 4 . Here, nine host biomarkers showed major concentration differences between the NP subjects and DWP individuals, both GP and LP. MMP-8, MMP-9, TIMP-1, and MIP-1α were the only markers that significantly differed between health and the subgroups of gingivitis (DWoP) patients. Resistin concentrations were significantly increased in the DWP periodontitis subgroups, while differences in adiponectin levels between disease categories were not observed. As the oral microbiome both drives and responds to host responses in the oral cavity, it was of interest to explore the relationship of targeted salivary biomolecules to the microbiome components and how that relationship was affected by diabetes and periodontal characteristics. Figure 5 A shows the frequency of significant correlations of the individual biomarker levels with the relative abundance of the 104 OTUs in the microbiome samples. These correlations were generally higher in the NP and DWP groups, with MMP-8, BAFF, adiponectin, and resistin showing the greatest frequency of positive correlations within the 3 subject groups. In contrast, there were only a limited number of negative correlations with any of the host molecules and the array of microbial taxa identified. Figure 5 B summarizes the details of the bacterial species that dominated these correlations with the 11 salivary biomarkers. As was expected few correlations with individual bacterial species were noted in the DWoP samples. The findings depicts 27 of the OTUs with extensive correlations to the salivary analytes, with 17 in the DWP patients and 10 in the NP subjects. The identified OTUs encompassed members of the Bacteroidetes, Firmicutes (Bacillota), Fusobacterium, Spirochaetes , and Synergistetes phyla. Supplemental Fig. 2A-D summarizes features of the correlations between individual OTUs of the 104 dominant members and levels of the individual salivary analytes. In the DWoP LG group, several significant correlations were observed, with the majority of positive correlations occurring with resistin and TIMP-1 (Fig. 2 A). In the DWoP GG patients, IL-1ß and adiponectin showed parallel patterns of positive correlations (Fig. 2 B). Additionally, correlations with MMP-8 and IL-6 were all positive but with different OTUs. All significant correlations with MIP-1α were negative in this patient subset. Figure 2 C summarizes the values for the DWP LP group that showed IFNα, TIMP-1, and adiponectin positively correlated with multiple OTUs. Finally, the largest frequency of correlations was noted in the DWP GP (Fig. 2 D) subjects. Here IL-1β, IL-6, IFNα, BAFF, adiponectin, and resistin were the predominant biomarkers that correlated with specific OTU, and frequently the same OTU. These various relationships allowed us to explore whether a targeted array of microbes and salivary biomarkers might be predictive of generalized and localized disease in T2DM patients. Odds Ratios (OR) were determined comparing levels of bacteria and host response biomolecules in periodontitis versus gingivitis subjects (Fig. 6 A). Here we observed OR of > 2.5 for most of the host biomarkers in the GP group, as well as increased ORs in the LP group. A similar number of oral bacteria exhibited ORs > 2 in LP or GP when compared to the gingivitis group. However, striking was an array of the bacterial species that displayed substantially lower ORs in the GP group compared to GG group. Figure 6 B provides a similar analysis for generalized versus localized inflammation or disease. Over 60% of the targeted host analytes showed ORs > 2 in LP compared with GP. Additionally, multiple bacterial species were specifically elevated in samples in the GP patients (ORs > 2). Similarly, multiple species of bacteria were specifically increased in the GG versus LG patients that were frequently different from those elevated in GP (eg. R. mucilaginosa, Prevotella sp. HMT305, V. dispar ). Also, a limited number of host response markers varied in the GG compared to the LG group (i.e., IL-6, MMP-8, adiponectin). DISCUSSION Epidemiological studies confirm that diabetes is a significant risk factor for periodontitis, and periodontitis increases the risk of poor diabetes management and sequelae in affected individuals. 16,39-42 Thus, this interplay between these chronic inflammatory conditions reflects the difficulty in controlling glucose levels and the initiation and progression of periodontitis. 43-49 The related clinical features reflect dysregulated inflammatory responses that are linked to tissue changes in T2DM, a microbiome contribution from periodontitis, and genetic regulation of the inflammatory status of the individual. l 43,50-55 Extensive reports identify the bidirectional linkage of diabetes and periodontitis, and studies that identify biological factors that contribute to the various stages of periodontal disease are not well characterized with these comorbidities. Hence, we sought to further characterize the salivary microbiome and associated host responses in T2DM patients (i.e., T2DM patients with periodontitis [DWP] or with gingivitis [DWoP]), with the goal of better understanding the panel of biological factors involved in the different extent and categories of periodontal disease compared to a NP healthy control group. This report thus described differences in the microbiomes at the individual patient level regarding the phyla distribution, and significant differences between localized and generalized gingivitis and periodontitis. Of the 104 dominant OTUs, approximately ¼ of these bacterial identifiers showed significant differences between health and the various disease presentation of localized and generalized periodontal diseases. These findings are consistent with the pathogenic microbiome in periodontitis, and particularly with more generalized disease. Considerable variation in inflammatory biomolecules concentrations have been observed in gingival crevicular fluid, saliva, and blood of patients with diabetes and periodontal disease. 56-60 Other studies have attempted to associate these mediators with periodontitis and the transition from health to metabolic syndrome and even T2DM. 61 Substantial evidence attributes pathogenic mechanisms linking periodontitis and diabetes to dysregulated inflammation. 62 We have shown previously in this population differences in salivary biomarkers between normal subjects and patients with T2DM, with or without periodontitis 29 that was consistent with the extant literature. However, in spite of the large number of studies on inflammatory mechanisms in periodontitis and diabetes, few have documented oral microbiome features that relate with specific host responses in the presence of these comorbidities. Various classic periodontal pathogens have been identified in diabetic and non-diabetic periodontitis patients, with P. gingivalis appearing in higher levels in both T2DM and T1DM patients. 63-67 A growing body of evidence suggests that diabetes may alter the local periodontal pocket environment favoring certain bacterial species to emerge. However, existing studies suggest some subtle differences in the oral microbiomes in diabetic patients, but clear clinical relevance of these differences have not been discerned. In the present study, significant differences were observed in various salivary analytes, especially MMP-8, MMP-9, TIMP-1, IL-1b, BAFF, and resistin primarily in the DWP versus both DWoP and NP subjects. Exploration of the relationship between the salivary analytes and individual members of the oral microbiome showed a greater number of significant, generally positive, correlations with MMP-8, BAFF, IFNa, adiponectin, and resistin in the DWP individuals. As the relationship between the chronic diseases of T2DM and periodontitis have emphasized a dysregulated host response, these findings support alterations in the host-microbe interactions when these diseases are coincident. Our previous report described differences in the oral microbiomes of normal subjects, T2DM patients without periodontitis (DWoP), and T2DM patients with periodontitis (DWP) (Ebersole et al., in review). In this analysis, we also identified several differences in the microbiomes within the DWoP and DWP groups that were stratified based upon localized or generalized clinical presentation of inflammation and destructive disease. While members of the Firmicutes (Bacillota) phyla were the dominant group of bacteria in both DWP and DWoP patients, there were clear differences in the GG (DWoP) and GP (DWP) subgroups showing elevated abundance of Bacteroidetes, Fusobacteria, Spirochaetes, and Synergistetes compared to the localized disease group. Thus, for the first time we demonstrated within the T2DM stratification, microbiomic variations appeared to relate to the magnitude (extent, severity) of oral inflammation and disease. For oral health in systemically healthy subjects or T2DM patients, there is a critical balance between tissue homeostasis and a transition into the microbial driven disease of periodontitis. Both genetic and environmental features regulate features of an individual’s response to the burden and quality of the oral microbiome. 42,68-70 As we have shown previously, there are distinctive differences in targeted host response biomolecules in saliva of these T2DM patients, as well as the characteristics of the salivary microbiome patterns coincident with these altered host response profiles. 33 We now identified clear differences in both the microbiome and salivary analytes related to disease extent in T2DM patients. Additionally, correlations between selected salivary analytes and specific members of the microbiome appeared to vary with disease extent, with the T2DM patients who had GP showing the greatest number of significant relationships. This type of analysis does not provide any cause-and-effect understanding of the processes that occur to reach this disease susceptible milieu in the oral environment. However, the results appear to reflect certain bacterial genera/species including Actinomyces _unclassified, Pr. dentalis, F. nucleatum _ssp._ vincentii, Leptotrichia sp._HMT_218, P. endodontalis, Sneathia _unclassified, T. denticola , and M. faucium that relate more directly to specific salivary analytes, such as IL-6, adiponectin, BAFF, and resistin. Consideration of these host and microbial biomarkers could be useful in portending the likelihood of more severe disease and aid in improved clinical decision-making on patient specific therapy. Limitations of this study include an overall number of subjects that were stratified based upon periodontal disease clinical characteristics of health or extent of oral disease that reduced the individual subgroup size. As such, the findings observed from these small subgroups may not be biologically generalizable to the broader T2DM or non-T2DM population. Also, poorly controlled diabetics were not included and may manifest distinctive host and microbiome traits. 50,71 This study, as most oral microbiome reports, describe the microbial ecology based upon a discrete sample thus lacking a general assessment of the permanency or transitory nature of the salivary microbiome in individual subjects. Finally, although saliva reflected many microbiome patterns apparent in the subgingival environment, saliva may not reflect all aspects important for understanding the biology of periodontitis. METHODS Study and Participants This cohort clinical study, approved by the Institutional Review Board of the University of Kentucky (UK) #17-0439-F6A in accordance with the Helsinki Declaration, was performed from June 2017 to July 2019. Inclusion criteria were: 35-80 years, a minimum of 16 teeth (excluding 3rd molars), in good systemic health (excluding the case definition), or diagnosed with T2DM based on the criteria of the American Diabetes Association. 28 We have reported previously that T2DM was confirmed by the patient's physician with glycated hemoglobin A1c (HbA1c), body mass index (BMI), and complete head, neck, and oral examination were recorded 29 . All subjects provided signed informed consent reviewed and accepted by the University of Kentucky Institutional Review Board. A full-mouth periodontal examination was conducted by a single calibrated dental examiner. Periodontal health measures included bleeding on probing (BOP), pocket depth (PD) and clinical attachment loss (CAL) at six sites per tooth. 30 Three cohorts were enrolled: (a) T2DM subjects with chronic periodontitis (DWP; n = 29), (b) T2DM subjects without chronic periodontitis (DWoP; n = 32) and (c) a cohort of healthier participants designated as ‘Not-Periodontitis’ (NP) (n = 31). Periodontitis was defined as having at least four teeth in two quadrants with a probing depth (PD) of ≥5 mm, clinical attachment loss (AL) of ≥2 mm and bleeding upon probing (BOP). 31,32 Participants in the NP group were defined as (a) no diabetes, (b) 1 (pinpoint bleeding on gentle probing), (c) 5 mm, (e) 2 mm with concurrent BOP and PD of > 4 mm. The three groups were balanced for age and sex. The extent of gingivitis and periodontitis in each patient was evaluated and used to categorize the patients as displaying localized / generalized gingivitis (LG, GG) or localized / generalized periodontitis (LP, GP). The DWoP group demonstrated gingivitis (i.e., BOP at 20% BOP sites [GG] coupled with all having less than five sites with > 4 mm periodontal pockets). The DWP group demonstrated either LP (4-87% BOP; 4 mm pockets) or GP (21-97% BOP; >10% sites with > 4 mm pockets). As reported previously 33 , medical and dental histories were obtained and subjects were excluded with alcoholism; liver, kidney or salivary gland dysfunction; inflammatory bowel disease; granulomatous diseases; immunosuppressive or cancer therapy. Additionally, acute illness ( i.e., fever, sore throat, body aches and diarrhea), pregnancy or lactation, use of antibiotics within the last 6 months, need for antibiotics for dental procedures, or the presence of an oral mucosal inflammatory conditions (e.g., aphthous, lichen planus, leukoplakia, and oral cancer) and current smoker were exclusion criteria. Biologic samples and analysis Unstimulated whole saliva samples was obtained from each participant, managed, stored, and concentrations of interleukin (IL)-1β, interleukin-6 (IL-6), MIP-1a (macrophage inflammatory protein 1 alpha), BAFF (B-cell activating factor; TNFSF13b), IFNa (interferon alpha), adiponectin, and resistin were measured in duplicate using Luminex technology with human cytokine/chemokine multiplex kits (Millipore, St. Charles, MO, USA). Salivary concentrations of matrix metalloprotease (MMP)-8, MMP-9 and TIMP-1 (tissue inhibitor of matrix matellaoproteinases-1) were determined in duplicate for each subject using human Quantikine enzyme-linked immunosorbent assay kits (R&D Systems, Minneapolis, MN, USA, as we have previously described. 29,34,35 Salivary microbiome characteristics were determined in each subject’s samples using 16S rRNA sequencing as we have previously described. 36-38 Sequences were assigned to their respective taxonomic classification using the Human Oral Microbiome Database (HOMD V13) (http://www.homd.org/index.php?name=seqDownload&file&type=R). The raw data are deposited at BioProject ID PRJNA516659 through the NIH NCBI. Statistical Analysis Descriptive statistics were performed for the clinical features, Operational Taxonomic Unit (OTU) microbiome differences, and log transformed salivary analyte comparisons through ANOVA or two sample t-test. Tukey’ honest significant difference method (HSD) was used for pairwise comparisons of the variables among groups of patients, if an ANOVA test was significant. The statistical software package (SAS 9.4, Cary, NC; IBM Inc., 2020) was used for the analysis and the statistical significance level was set at 0.05. Correlation analyses were evaluated using a Pearson correlation coefficient with a significant level of 0.05. The data for gene expression have been uploaded into GEO accession GSE180588 (https://www.ncbi.nlm.nih.gov/gds). Declarations ACKNOWLEDGEMENTS We thank Sabrina Braden, Dawn Dawson and Jason Stevens for their contributions in the recruitment, enrollment, data collection and laboratory analyses. This study was supported by the Delta Dental Foundation of Michigan. Author Contributions Author contributions include JLE and CSM contributed to conception, design, data acquisition and interpretation, drafted and critically revised the manuscript. SSK contributed to data acquisition, analysis, and interpretation, and critically reviewed the manuscript. DDIII contributed to obtaining and interpreting the clinical data and critically reviewed the manuscript. XDZ provided biostatistical evaluation for data analysis, interpretation, and critical revision of the manuscript. All authors gave their final approval and agreement to be accountable for all aspects of the work. The authors state no conflict with any information provided in the report. Data Availability The microbiome data are deposited at BioProject ID PRJNA516659 through the NIH NCBI. The data for gene expression have been uploaded into GEO accession GSE180588 (https://www.ncbi.nlm.nih.gov/gds). References Rohm, T. V., Meier, D. T., Olefsky, J. M. & Donath, M. Y. Inflammation in obesity, diabetes, and related disorders. Immunity 55, 31–55, doi: 10.1016/j.immuni.2021.12.013 (2022). Geisinger, M. L. et al. Systemic Inflammatory Biomarkers and Their Association With Periodontal and Diabetes-Related Factors in the Diabetes and Periodontal Therapy Trial, A Randomized Controlled Trial. J Periodontol 87, 900–913, doi: 10.1902/jop.2016.150727 (2016). Girard, D. & Vandiedonck, C. How dysregulation of the immune system promotes diabetes mellitus and cardiovascular risk complications. Front Cardiovasc Med 9, 991716, doi: 10.3389/fcvm.2022.991716 (2022). Velikova, T. V., Kabakchieva, P. P., Assyov, Y. S. & Georgiev Tcapital A, C. Targeting Inflammatory Cytokines to Improve Type 2 Diabetes Control. BioMed research international 2021, 7297419, doi: 10.1155/2021/7297419 (2021). Prattichizzo, F. et al. Inflammageing and metaflammation: The yin and yang of type 2 diabetes. Ageing research reviews 41, 1–17, doi: 10.1016/j.arr.2017.10.003 (2017). Gokhale, N. H. et al. Resistin levels in gingival crevicular fluid of patients with chronic periodontitis and type 2 diabetes mellitus. J Periodontol 85, 610–617, doi: 10.1902/jop.2013.130092 (2014). Mohan, M. et al. Impact of scaling and root planing on C-reactive protein levels in gingival crevicular fluid and serum in chronic periodontitis patients with or without diabetes mellitus. J Periodontal Implant Sci 44, 158–168, doi: 10.5051/jpis.2014.44.4.158 (2014). Poston, C. J. et al. Statin intake is associated with MMP-1 level in gingival crevicular fluid of patients with periodontitis. Oral Dis 22, 438–444, doi: 10.1111/odi.12474 (2016). Techatanawat, S. et al. Salivary and serum interleukin-17A and interleukin-18 levels in patients with type 2 diabetes mellitus with and without periodontitis. PLoS One 15, e0228921, doi: 10.1371/journal.pone.0228921 (2020). Hirtz, C. et al. The potential impact of salivary peptides in periodontitis. Crit Rev Clin Lab Sci 58, 479–492, doi: 10.1080/10408363.2021.1907298 (2021). Nibali, L. et al. Patterns of subgingival microbiota in different periodontal phenotypes. J Dent 117, 103912, doi: 10.1016/j.jdent.2021.103912 (2022). Johnston, W. et al. Mechanical biofilm disruption causes microbial and immunological shifts in periodontitis patients. Scientific reports 11, 9796, doi: 10.1038/s41598-021-89002-z (2021). Zhang, S., Yu, N. & Arce, R. M. Periodontal inflammation: Integrating genes and dysbiosis. Periodontol 2000 82, 129–142, doi: 10.1111/prd.12267 (2020). Na, H. S. et al. Identification of Potential Oral Microbial Biomarkers for the Diagnosis of Periodontitis. J Clin Med 9, doi: 10.3390/jcm9051549 (2020). Curtis, M. A., Diaz, P. I. & Van Dyke, T. E. The role of the microbiota in periodontal disease. Periodontol 2000 83, 14–25, doi: 10.1111/prd.12296 (2020). Polak, D., Sanui, T., Nishimura, F. & Shapira, L. Diabetes as a risk factor for periodontal disease-plausible mechanisms. Periodontol 2000 83, 46–58, doi: 10.1111/prd.12298 (2020). Sudhakara, P., Gupta, A., Bhardwaj, A. & Wilson, A. Oral Dysbiotic Communities and Their Implications in Systemic Diseases. Dent J (Basel) 6, 10, doi: 10.3390/dj6020010 (2018). Sabharwal, A. et al. The salivary microbiome of diabetic and non-diabetic adults with periodontal disease. J Periodontol, doi: 10.1002/JPER.18-0167 (2018). Pascale, A. et al. Microbiota and metabolic diseases. Endocrine 61, 357–371, doi: 10.1007/s12020-018-1605-5 (2018). Nibali, L. et al. Differences in the periodontal microbiome of successfully treated and persistent aggressive periodontitis. J Clin Periodontol 47, 980–990, doi: 10.1111/jcpe.13330 (2020). Lu, H. et al. Well-maintained patients with a history of periodontitis still harbor a more dysbiotic microbiome than health. J Periodontol 91, 1584–1594, doi: 10.1002/JPER.19-0498 (2020). Liu, G. et al. Shift in the subgingival microbiome following scaling and root planing in generalized aggressive periodontitis. J Clin Periodontol 45, 440–452, doi: 10.1111/jcpe.12862 (2018). Simpson, T. C. et al. Treatment of periodontitis for glycaemic control in people with diabetes mellitus. The Cochrane database of systematic reviews 4, CD004714, doi: 10.1002/14651858.CD004714.pub4 (2022). Jain, P. et al. Periodontitis and Systemic Disorder-An Overview of Relation and Novel Treatment Modalities. Pharmaceutics 13, doi: 10.3390/pharmaceutics13081175 (2021). Kapellas, K. et al. Periodontal therapy and glycaemic control among individuals with type 2 diabetes: reflections from the PerioCardio study. International journal of dental hygiene, doi: 10.1111/idh.12234 (2016). Michalowicz, B. S. et al. Factors associated with the clinical response to nonsurgical periodontal therapy in people with type 2 diabetes mellitus. J Am Dent Assoc 145, 1227–1239, doi: 10.14219/jada.2014.92 (2014). Gaikwad, S. P., Gurav, A. N., Shete, A. R. & Desarda, H. M. Effect of scaling and root planing combined with systemic doxycycline therapy on glycemic control in diabetes mellitus subjects with chronic generalized periodontitis: a clinical study. J Periodontal Implant Sci 43, 79–86, doi: 10.5051/jpis.2013.43.2.79 (2013). American, D. A. Diagnosis , (2022). Miller, C. S., Ding, X., Dawson, D. R., 3rd & Ebersole, J. L. Salivary biomarkers for discriminating periodontitis in the presence of diabetes. J Clin Periodontol 48, 216–225, doi: 10.1111/jcpe.13393 (2021). Dawson, D. R., 3rd et al. Salivary levels of Epstein-Barr virus DNA correlate with subgingival levels, not severity of periodontitis. Oral Dis 15, 554–559, doi: 10.1111/j.1601-0825.2009.01585.x (2009). Armitage, G. C. Development of a classification system for periodontal diseases and conditions. Annals of periodontology / the American Academy of Periodontology 4, 1–6, doi: 10.1902/annals.1999.4.1.1 (1999). Armitage, G. C., Research, S. & Therapy Committee of the American Academy of, P. Diagnosis of periodontal diseases. J Periodontol 74, 1237–1247, doi: 10.1902/jop.2003.74.8.1237 (2003). Miller, C. S., Ding, X., Nagarajan, R., Dawson, D. R., 3rd & Ebersole, J. L. Biomarker panel discriminates diabetics with and without periodontitis pre- and post-therapy. J Periodontal Res 58, 493–502, doi: 10.1111/jre.13127 (2023). Nagarajan, R., Miller, C. S., Dawson, D., 3rd, Al-Sabbagh, M. & Ebersole, J. L. Patient-Specific Variations in Biomarkers across Gingivitis and Periodontitis. PLoS One 10, e0136792, doi: 10.1371/journal.pone.0136792 (2015). Ebersole, J. L., Nagarajan, R., Akers, D. & Miller, C. S. Targeted salivary biomarkers for discrimination of periodontal health and disease(s). Frontiers in cellular and infection microbiology 5, 62, doi: 10.3389/fcimb.2015.00062 (2015). Kirakodu, S., Chen, J., Gonzalez Martinez, J., Gonzalez, O. A. & Ebersole, J. Microbiome Profiles of Ligature-Induced Periodontitis in Nonhuman Primates across the Life Span. Infect Immun 87, e00067-00019, doi: 10.1128/IAI.00067-19 (2019). Kirakodu, S. S., Govindaswami, M., Novak, M. J., Ebersole, J. L. & Novak, K. F. Optimizing qPCR for the Quantification of Periodontal Pathogens in a Complex Plaque Biofilm. Open Dent J 2, 49–55, doi: 10.2174/1874210600802010049 (2008). Ebersole, J. L., Kirakodu, S. S., Dawson III, D., Nagarajan, R. & Miller, C. S. Microbiome characteristics of diabetics with periodontitis before and after treatment. Molecular Oral MIcrobiology in review (2024). Pussinen, P. J. et al. Periodontitis and cardiometabolic disorders: The role of lipopolysaccharide and endotoxemia. Periodontol 2000 89, 19–40, doi: 10.1111/prd.12433 (2022). Bhuyan, R. et al. Periodontitis and Its Inflammatory Changes Linked to Various Systemic Diseases: A Review of Its Underlying Mechanisms. Biomedicines 10, 2659, doi: 10.3390/biomedicines10102659 (2022). Bains, V. K., Mahendra, J., Mahendra, L., Mittal, M. & Valli, G. Markers, Pathways, and Current Evidence for Periodontitis-associated Insulin Resistance: A Narrative Review. Journal of International Society of Preventive & Community Dentistry 12, 475–487, doi: 10.4103/jispcd.JISPCD_92_22 (2022). Kapila, Y. L. Oral health's inextricable connection to systemic health: Special populations bring to bear multimodal relationships and factors connecting periodontal disease to systemic diseases and conditions. Periodontol 2000 87, 11–16, doi: 10.1111/prd.12398 (2021). Nascimento, G. G., Leite, F. R. M., Vestergaard, P., Scheutz, F. & Lopez, R. Does diabetes increase the risk of periodontitis? A systematic review and meta-regression analysis of longitudinal prospective studies. Acta Diabetol 55, 653–667, doi: 10.1007/s00592-018-1120-4 (2018). Genco, R. J. & Borgnakke, W. S. Risk factors for periodontal disease. Periodontol 2000 62, 59–94, doi: 10.1111/j.1600-0757.2012.00457.x (2013). Lalla, E. et al. Periodontal Changes in Children and Adolescents With Diabetes: A case-control study. Diabetes Care 29, 295–299 (2006). Demmer, R. T., Jacobs, D. R., Jr. & Desvarieux, M. Periodontal disease and incident type 2 diabetes: results from the First National Health and Nutrition Examination Survey and its epidemiologic follow-up study. Diabetes Care 31, 1373–1379, doi: 10.2337/dc08-0026 (2008). Borgnakke, W. S., Ylostalo, P. V., Taylor, G. W. & Genco, R. J. Effect of periodontal disease on diabetes: systematic review of epidemiologic observational evidence. J Clin Periodontol 40 Suppl 14, S135-152, doi: 10.1111/jcpe.12080 (2013). Albert, D. A. et al. Diabetes and oral disease: implications for health professionals. Ann N Y Acad Sci 1255, 1–15, doi: 10.1111/j.1749-6632.2011.06460.x (2012). Chapple, I. L., Genco, R. & working group 2 of the joint, E. F. P. A. A. P. w. Diabetes and periodontal diseases: consensus report of the Joint EFP/AAP Workshop on Periodontitis and Systemic Diseases. J Periodontol 84, S106-112, doi: 10.1902/jop.2013.1340011 (2013). Sanz, M. et al. Scientific evidence on the links between periodontal diseases and diabetes: Consensus report and guidelines of the joint workshop on periodontal diseases and diabetes by the International Diabetes Federation and the European Federation of Periodontology. J Clin Periodontol 45, 138–149, doi: 10.1111/jcpe.12808 (2018). Nguyen, A. T. M. et al. The association of periodontal disease with the complications of diabetes mellitus. A systematic review. Diabetes Res Clin Pract 165, 108244, doi: 10.1016/j.diabres.2020.108244 (2020). Graziani, F., Gennai, S., Solini, A. & Petrini, M. A systematic review and meta-analysis of epidemiologic observational evidence on the effect of periodontitis on diabetes An update of the EFP-AAP review. J Clin Periodontol 45, 167–187, doi: 10.1111/jcpe.12837 (2018). Ziukaite, L., Slot, D. E., Cobb, C. M., Coucke, W. & Van der Weijden, G. A. Prevalence of diabetes among patients diagnosed with periodontitis: A retrospective cross-sectional study. International journal of dental hygiene 16, 305–311, doi: 10.1111/idh.12280 (2018). Chapple, I. L. et al. Interaction of lifestyle, behaviour or systemic diseases with dental caries and periodontal diseases: consensus report of group 2 of the joint EFP/ORCA workshop on the boundaries between caries and periodontal diseases. J Clin Periodontol 44 Suppl 18, S39-S51, doi: 10.1111/jcpe.12685 (2017). Polak, D. & Shapira, L. An update on the evidence for pathogenic mechanisms that may link periodontitis and diabetes. J Clin Periodontol 45, 150–166, doi: 10.1111/jcpe.12803 (2018). Kardesler, L., Buduneli, N., Cetinkalp, S. & Kinane, D. F. Adipokines and inflammatory mediators after initial periodontal treatment in patients with type 2 diabetes and chronic periodontitis. J Periodontol 81, 24–33, doi: 10.1902/jop.2009.090267 (2010). Kardesler, L., Buduneli, N., Cetinkalp, S., Lappin, D. & Kinane, D. F. Gingival crevicular fluid IL-6, tPA, PAI-2, albumin levels following initial periodontal treatment in chronic periodontitis patients with or without type 2 diabetes. Inflamm Res 60, 143–151, doi: 10.1007/s00011-010-0248-7 (2011). Kardesler, L. et al. Crevicular fluid matrix metalloproteinase-8, -13, and TIMP-1 levels in type 2 diabetics. Oral Dis 16, 476–481, doi: 10.1111/j.1601-0825.2010.01659.x (2010). Navarro-Sanchez, A. B., Faria-Almeida, R. & Bascones-Martinez, A. Effect of non-surgical periodontal therapy on clinical and immunological response and glycaemic control in type 2 diabetic patients with moderate periodontitis. J Clin Periodontol 34, 835–843, doi: 10.1111/j.1600-051X.2007.01127.x (2007). Santos, V. R. et al. Cytokine levels in sites of chronic periodontitis of poorly controlled and well-controlled type 2 diabetic subjects. J Clin Periodontol 37, 1049–1058, doi: 10.1111/j.1600-051X.2010.01624.x (2010). Genco, R. J., Graziani, F. & Hasturk, H. Effects of periodontal disease on glycemic control, complications, and incidence of diabetes mellitus. Periodontol 2000 83, 59–65, doi: 10.1111/prd.12271 (2020). Taylor, J. J., Preshaw, P. M. & Lalla, E. A review of the evidence for pathogenic mechanisms that may link periodontitis and diabetes. J Periodontol 84, S113–134, doi: 10.1902/jop.2013.134005 (2013). Liu, L. S., Gkranias, N., Farias, B., Spratt, D. & Donos, N. Differences in the subgingival microbial population of chronic periodontitis in subjects with and without type 2 diabetes mellitus-a systematic review. Clin Oral Investig 22, 2743–2762, doi: 10.1007/s00784-018-2660-2 (2018). Lira-Junior, R., Akerman, S., Klinge, B., Bostrom, E. A. & Gustafsson, A. Salivary microbial profiles in relation to age, periodontal, and systemic diseases. PLoS One 13, e0189374, doi: 10.1371/journal.pone.0189374 (2018). Field, C. A., Gidley, M. D., Preshaw, P. M. & Jakubovics, N. Investigation and quantification of key periodontal pathogens in patients with type 2 diabetes. J Periodontal Res 47, 470–478, doi: 10.1111/j.1600-0765.2011.01455.x (2012). Makiura, N. et al. Relationship of Porphyromonas gingivalis with glycemic level in patients with type 2 diabetes following periodontal treatment. Oral Microbiol Immunol 23, 348–351, doi: 10.1111/j.1399-302X.2007.00426.x (2008). Ebersole, J. L., Holt, S. C., Hansard, R. & Novak, M. J. Microbiologic and immunologic characteristics of periodontal disease in Hispanic americans with type 2 diabetes. J Periodontol 79, 637–646, doi: 10.1902/jop.2008.070455 (2008). Loos, B. G. & Van Dyke, T. E. The role of inflammation and genetics in periodontal disease. Periodontol 2000 83, 26–39, doi: 10.1111/prd.12297 (2020). Dommisch, H., Kuzmanova, D., Jonsson, D., Grant, M. & Chapple, I. Effect of micronutrient malnutrition on periodontal disease and periodontal therapy. Periodontol 2000 78, 129–153, doi: 10.1111/prd.12233 (2018). Reynolds, M. A. Modifiable risk factors in periodontitis: at the intersection of aging and disease. Periodontol 2000 64, 7–19, doi: 10.1111/prd.12047 (2014). Miranda, T. S. et al. Influence of glycemic control on the levels of subgingival periodontal pathogens in patients with generalized chronic periodontitis and type 2 diabetes. Journal of applied oral science: revista FOB 25, 82–89, doi: 10.1590/1678-77572016-0302 (2017). Additional Declarations No competing interests reported. Supplementary Files Deltadentalhostsupplemental524.pptx Supplemental Figure 1: Summation of total reads for the detected OTUs in the saliva samples from various subsets of subjects. Each point denotes the value for the sample from one subject. The horizontal bar denotes the mean reads for that group. Supplemental Figure 2: Correlations for individual microbiome OTUs and the salivary analytes in T2DM patients with generalized (A; GG) and localized (B; LG) gingivitis, as well as generalized (C; GP), and localized (D; LP) periodontitis. The points denote the correlation coefficient for a specific OTU as shown on the X-axis (see Table 1). The point above or below the shaded rectangles distinguish the salivary analyte and associated OTU that exceeded significant positive or negative correlation values (p<0.01). Cite Share Download PDF Status: Published Journal Publication published 28 Dec, 2024 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 15 Jul, 2024 Reviews received at journal 14 Jul, 2024 Reviewers agreed at journal 14 Jul, 2024 Reviews received at journal 14 Jul, 2024 Reviewers agreed at journal 03 Jul, 2024 Reviews received at journal 05 Jun, 2024 Reviewers agreed at journal 31 May, 2024 Reviewers agreed at journal 31 May, 2024 Reviewers invited by journal 31 May, 2024 Editor assigned by journal 31 May, 2024 Editor invited by journal 31 May, 2024 Submission checks completed at journal 31 May, 2024 First submitted to journal 08 May, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4391317","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":312640157,"identity":"18a2f8bc-16a1-48c7-80c3-62b5fe2feae5","order_by":0,"name":"Jeffrey L. 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The DWoP group had localized (LG) or generalized (GG) gingivitis. The DWP group had localized (LP) or generalized (GP) periodontitis. The bars are the group means and the vertical brackets enclose 1 SEM. The connecting horizontal bars identify significant differences between the groups and the asterisk (*) for BMI values denotes significantly different than the NP group at \u003cem\u003ep\u003c/em\u003e\u0026lt;0.05 as determined by an ANOVA.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4391317/v1/0e7c604bc1903cf1beb3fc61.png"},{"id":58385276,"identity":"942b35fd-3735-4232-bd56-cc2faa098b87","added_by":"auto","created_at":"2024-06-14 18:39:19","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":544694,"visible":true,"origin":"","legend":"\u003cp\u003eRelative abundance of different phyla to total sequence readouts in individual (\u003cstrong\u003eA\u003c/strong\u003e) NP, (\u003cstrong\u003eB\u003c/strong\u003e) DWoP, or (\u003cstrong\u003eC\u003c/strong\u003e) DWP subjects identified as generalized or localized disease. Panel (a) and (b) stratify based upon the relative abundance of the phyla. The table provides mean ± SD of relative abundance of each phylum across the subject group.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4391317/v1/202857b8f856f260b0b37676.png"},{"id":58385275,"identity":"2aa153ea-958e-4a08-b1ae-f9c9096649bf","added_by":"auto","created_at":"2024-06-14 18:39:19","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":119341,"visible":true,"origin":"","legend":"\u003cp\u003eRelative abundance of OTUs (eg. species, genera) in to total sequence readouts of NP, DWoP (GG, LG), and DWP (GP, LP) in salivary microbiome samples. Panels A, B and C organize the microbes based upon general relative abundance across the samples. Bars denote group means with asterisk (*) signifying a significant difference between NP and disease groups, and hashtag (#) identifying one or more disease subsets that differs in OTU (eg. species, genera) abundance from the other groups. OTUs were only included if a significant difference at p\u0026lt;0.01 was detected for any comparison as determined by ANOVA.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4391317/v1/e5a8a508efa3ca81d360c071.png"},{"id":58387136,"identity":"d15b8826-1116-4f77-a6c2-c2ab5cb49d8f","added_by":"auto","created_at":"2024-06-14 18:47:19","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":95404,"visible":true,"origin":"","legend":"\u003cp\u003eLevels of salivary analytes in subsets of DWoP and DWP T2DM patients with localized (LG, LP) or generalized (GG, GP) disease compared to control subjects (NP). Bars denote group means and vertical brackets signify 1 SEM. Significance is indicated compared to the NP group, as well as between the various disease strata as determined by ANOVA.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4391317/v1/bfbec702bdaeeee209aa13f4.png"},{"id":58385278,"identity":"d779ca57-09fb-44c3-a870-83c819e89b5d","added_by":"auto","created_at":"2024-06-14 18:39:19","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":174454,"visible":true,"origin":"","legend":"\u003cp\u003eFrequency of OTUs with significant correlations with levels of the individual salivary analytes. (\u003cstrong\u003eA\u003c/strong\u003e) Bars denote the number of positive or negative correlations for each analyte within the groups. (\u003cstrong\u003eB\u003c/strong\u003e) Frequency of significant correlations across the 11 salivary analytes with the individual OTUs. Each dot represents the number of significant (\u003cem\u003ep\u003c/em\u003e\u0026lt;0.05) correlations for a specific subject group. Only OTUs were included where there was any significant correlation in one or more patient groups.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-4391317/v1/ebe5cc16d4d1008c679fab24.png"},{"id":58387694,"identity":"e0022cb7-b4a4-4070-8385-9f3832486b54","added_by":"auto","created_at":"2024-06-14 18:55:19","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":78890,"visible":true,"origin":"","legend":"\u003cp\u003eDisplay of Odd Ratios (OR) for various microbes and host factors comparing (\u003cstrong\u003eA\u003c/strong\u003e) localized gingivitis (LG) to localized periodontitis (LP) and generalized gingivitis (GG) to generalized periodontitis (GP), and (\u003cstrong\u003eB\u003c/strong\u003e) LG to GG and LP to GP in the T2DM groups. Each point denotes an OR value for the specific variable (microbe, host factor). Positive OR denotes elevated level in generalized disease compared to localized disease or in gingivitis vs. periodontitis. A negative OR denotes elevated level in LG, LP, or gingivitis compared to generalized disease or periodontitis, respectively.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-4391317/v1/fd4a5967664988fde662fa37.png"},{"id":72640844,"identity":"fb13f2e4-c383-4040-93f6-d1441d635b9b","added_by":"auto","created_at":"2024-12-30 16:10:20","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1614332,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4391317/v1/15d9dab6-6188-40f9-a7d8-6966fbd7694d.pdf"},{"id":58387135,"identity":"49b2f06f-24e9-4d73-86ed-84962b4dc67f","added_by":"auto","created_at":"2024-06-14 18:47:19","extension":"pptx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":301244,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplemental Figure 1:\u003c/strong\u003e Summation of total reads for the detected OTUs in the saliva samples from various subsets of subjects. Each point denotes the value for the sample from one subject. The horizontal bar denotes the mean reads for that group.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSupplemental Figure 2:\u003c/strong\u003e Correlations for individual microbiome OTUs and the salivary analytes in T2DM patients with generalized (\u003cstrong\u003eA\u003c/strong\u003e; GG) and localized (\u003cstrong\u003eB\u003c/strong\u003e; LG) gingivitis, as well as generalized (\u003cstrong\u003eC\u003c/strong\u003e; GP), and localized (\u003cstrong\u003eD\u003c/strong\u003e; LP) periodontitis. The points denote the correlation coefficient for a specific OTU as shown on the X-axis (see Table 1). The point above or below the shaded rectangles distinguish the salivary analyte and associated OTU that exceeded significant positive or negative correlation values (p\u0026lt;0.01).\u003c/p\u003e","description":"","filename":"Deltadentalhostsupplemental524.pptx","url":"https://assets-eu.researchsquare.com/files/rs-4391317/v1/200c6b3b88bdc9ec903fb49e.pptx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Salivary Features of Periodontitis and Gingivitis in Type 2 Diabetes Mellitus","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eType 2 diabetes mellitus (T2DM) is a chronic inflammatory disease with elevated systemic biomolecules of inflammation that are central to the pathogenesis of the disease.\u003csup\u003e\u003cspan additionalcitationids=\"CR2 CR3 CR4\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e Beyond the identification of periodontitis as a sequelae of T2DM, localized inflammatory proteins are routinely increased in the gingival crevicular fluid and saliva of periodontitis affected T2DM patients.\u003csup\u003e\u003cspan additionalcitationids=\"CR7 CR8 CR9\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e Within this oral environment the host responses are driven by, and reflect, an altered microbiome with dysbiotic changes of opportunistic pathogens and pathobionts revealing a lesional process.\u003csup\u003e\u003cspan additionalcitationids=\"CR12 CR13 CR14\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eAs variations in the characteristics of the oral microbiome and associated host responses have been related to extrinsic and intrinsic (e.g., T2DM, age) effectors of the host ecosystem \u003csup\u003e\u003cspan additionalcitationids=\"CR17 CR18\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e, there remain gaps in our knowledge concerning the biologic mechanisms that are controlling these features in the oral cavity. Additionally, mechanical therapeutics in systemically normal individuals have been shown to substantially impact the microbiome and help transition its characteristics to one more consistent with periodontal health.\u003csup\u003e\u003cspan additionalcitationids=\"CR21\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e Nevertheless, existing data indicate that the prevalence and severity of periodontitis is increased in T2DM\u003csup\u003e\u003cspan additionalcitationids=\"CR24 CR25 CR26\" citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThis investigation cross-tabulated the microbiome and biomarkers in saliva of T2DM patients with or without periodontitis. The findings provide insight into the unique features of localized and generalized gingivitis and periodontal disease in the presence of T2DM.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eOral clinical features in diabetes\u003c/h2\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e provides the periodontal characteristics and BMI relative to the various subgroups of T2DM and NP subjects. Only the NP group showed a significantly lower BMI compared to most of the T2DM groups (e.g., DWoP, DWP). In contrast and as expected, the clinical features of destructive disease in the generalized periodontitis patients were significantly different from both the gingivitis and localized periodontitis groups. Additionally, BOP showed significantly greater levels with increasing severity of disease. Of note, the majority of periodontally healthy individuals were in the NP group, and the majority of gingivitis cases were in the DWoP group. Based upon the disease features in affected patients, an additional perspective evaluated the characteristics of the microbiome and host response patterns related to the magnitude of disease.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eMicrobiome characteristics in generalized and localized disease in diabetes\u003c/h2\u003e \u003cp\u003e \u003cb\u003eSupplemental Fig.\u0026nbsp;1\u003c/b\u003e provides a summary of the individual sample OTU reads for the NP, GG, LG, GP, and LP salivary microbiomes. Generally, similar numbers of overall reads were identified across the subgroups. In another report we identified the top 104 OTUs based upon levels across the three study groups (NP, DWoP, DWP), and showed that this portfolio of microorganisms provide 96\u0026ndash;98% coverage of the microbiome reads in the samples \u003csup\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA presents the results for the individual NP subjects and shows \u003cem\u003eFirmicutes (Bacillota)\u003c/em\u003e as the dominant phyla, with the relative abundance varying from ~\u0026thinsp;40\u0026ndash;80%. \u003cem\u003eBacteroidetes\u003c/em\u003e phyla was next most prevalent, being detected in 23 subjects. \u003cem\u003eProteobacteria\u003c/em\u003e was observed in the remaining six individuals. A subset of NP subjects demonstrated elevated levels of \u003cem\u003eSpirochetes\u003c/em\u003e and four individuals had elevated \u003cem\u003eAbsconditabacteria\u003c/em\u003e in their salivary microbiome.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eOperational taxonomic units (OTUs) included in the analysis.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOTU\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePhyla\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGenera/Species\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu150\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAbsconditabacteria_(SR1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAbsconditabacteria_(SR1)_[G-1] bacterium_HMT_345\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu130\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAbsconditabacteria_(SR1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAbsconditabacteria_(SR1)_[G-1] bacterium_HMT_875\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu025\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eActinomyces graevenitzii\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eActinomyces sp._HMT_169\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu007\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eActinomyces sp._HMT_180\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eActinomyces_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu049\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAtopobium_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu106\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBifidobacterium longum\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu053\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCorynebacterium matruchotii\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRothia mucilaginosa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu080\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eActinobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRothia_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacillota\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSelenomonas artemidis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu163\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacillota\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSelenomonas sp._HMT_126\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacillota\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSelenomonas sp._HMT_136\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu034\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacillota\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSelenomonas sputigena\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu019\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacillota\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSelenomonas_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu048\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacillota\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStomatobaculum longum\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu087\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacillota\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStomatobaculum sp._HMT_097\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu064\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlloprevotella rava\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu058\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlloprevotella sp._HMT_308\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu111\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlloprevotella sp._HMT_473\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu042\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAlloprevotella tannerae\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu147\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBacteroidales_[G-2] bacterium_HMT_274\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu098\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBergeyella sp._HMT_322\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu062\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCapnocytophaga gingivalis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu063\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCapnocytophaga granulosa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu065\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCapnocytophaga leadbetteri\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu074\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCapnocytophaga sputigena\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu083\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePorphyromonas endodontalis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu057\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePorphyromonas gingivalis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu038\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePorphyromonas pasteri\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu173\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella dentalis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu054\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella denticola\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu136\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella intermedia\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu033\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella nanceiensis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu070\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella nigrescens\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu018\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella oris\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu068\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella oulorum\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu017\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella pallens\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu164\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella pleuritidis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu015\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella salivae\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu092\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella sp._HMT_300\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu073\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella sp._HMT_305\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu011\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella veroralis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePrevotella_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu085\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBacteroidetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTannerella sp._HMT_286\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu067\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCatonella morbi\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu135\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDialister invisus\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu031\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGemella morbillorum\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu027\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGranulicatella adiacens\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu056\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLachnoanaerobaculum orale\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLachnoanaerobaculum umeaense\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu035\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLachnospiraceae_[G-2] bacterium_HMT_096\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu022\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMegasphaera micronuciformis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu158\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMogibacterium vescum\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu252\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMycoplasma faucium\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu051\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOribacterium asaccharolyticum\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu052\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOribacterium_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu112\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eParvimonas micra\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu137\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePeptostreptococcaceae_[XI][G-1]_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu078\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePeptostreptococcus stomatis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu094\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRuminococcaceae_[G-1] bacterium_HMT_075\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu129\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRuminococcaceae_[G-2] bacterium_HMT_085\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStreptococcus oralis_subsp._dentisani_clade_058\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu008\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStreptococcus parasanguinis_clade_411\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu006\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStreptococcus salivarius\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStreptococcus_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVeillonella dispar\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu009\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVeillonella parvula\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu119\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVeillonella sp._HMT_780\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu069\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVeillonella sp._HMT_917\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu002\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVeillonella_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu152\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFirmicutes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eVeillonellaceae_[G-1] bacterium_HMT_155\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFusobacterium nucleatum_subsp._vincentii\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu059\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFusobacterium sp._HMT_203\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFusobacterium_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu044\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia hongkongensis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu047\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia sp._HMT_212\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu020\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia sp._HMT_215\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu095\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia sp._HMT_218\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu045\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia sp._HMT_221\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu185\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia sp._HMT_223\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu046\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia sp._HMT_392\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu055\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia wadei\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu016\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLeptotrichia_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu165\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFusobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSneathia_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu071\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCampylobacter rectus\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu171\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCampylobacter sp._HMT_044\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCampylobacter_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu091\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEscherichia coli\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu110\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHaemophilus influenzae\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHaemophilus_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu115\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eKingella_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu097\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLautropia mirabilis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu028\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNeisseria macacae\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu066\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNeisseria oralis\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu039\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNeisseria perflava\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu021\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNeisseria_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu050\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProteobacteria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePasteurellaceae_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu146\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpirochaetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTreponema denticola\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu099\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpirochaetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTreponema socranskii\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSynergistetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFretibacterium fastidiosum\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu123\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSynergistetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFretibacterium sp._HMT_361\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOtu114\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSynergistetes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFretibacterium_unclassified\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eWe next stratified the diabetic periodontitis (DWP) and gingivitis (DWoP) patients based on the clinical presentation (i.e., generalized or localized form) of periodontal inflammation and lesions to examine the phyla distribution. \u003cem\u003eFirmicutes\u003c/em\u003e was the dominant phyla across the majority of DWoP patients with mean levels similar between generalized and localized gingivitis (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Three individuals showed \u003cem\u003eBacteroidetes\u003c/em\u003e as the dominant phyla. Generally, this phylum appeared more prevalent in the GG patients, while the \u003cem\u003eProteobacteria\u003c/em\u003e phylum was elevated across the LG subset. An elevated abundance was seen in LG for \u003cem\u003eAbsconditabacteria.\u003c/em\u003e Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC presents the phyla data for the DWP group with GP and LP. These generalized disease adults showed higher levels of \u003cem\u003eBacteroidetes, Fusobacteria, Proteobacteria, Spirochaetes\u003c/em\u003e, and \u003cem\u003eSynergistetes\u003c/em\u003e compared to the localized disease group although the differences did not reach statistical significance.\u003c/p\u003e \u003cp\u003eThe evaluation of the microbiome differences was extended to focus on the individual microbial components in the various patient groups. Figure\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e illustrates the relative abundance of the targeted set of 104 OTUs in samples from the study population based on the level of oral disease (i.e., NP [healthy], LG, GG, LP, and GP). We identified 47 OTUs (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) that were significantly different in abundance between NP and any disease subset or between disease subgroup categories (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). There also were numerous OTU abundance differences (N\u0026thinsp;=\u0026thinsp;18) between LG and LP. Seven bacterial species differences in abundance were observed between GG and GP, four bacterial species abundance differences were observed between LG and GG, and only one species (\u003cem\u003eP. gingivalis\u003c/em\u003e) that was different in relative abundance between LP and GP.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMicrobiome relative abundance differences between generalized and localized disease in T2DM patients. These taxa are significantly different between the groups determined by a \u003cem\u003et test\u003c/em\u003e (see Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGG vs. LG\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGP vs. LP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGG vs. GP\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLG vs. LP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePrevotella\u003c/em\u003e_unclassified\u003c/p\u003e \u003cp\u003e\u003cem\u003eM. micronuciformis\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ePrevotella\u003c/em\u003e sp._HMT_305\u003c/p\u003e \u003cp\u003e\u003cem\u003eBergeyella\u003c/em\u003e sp._HMT_322\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eP. gingivalis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eF. nucleatum_subsp._vincentii\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ePrevotella nigrescens\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eRothia_unclassified\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eAlloprevotella tannerae\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eP. gingivalis\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eCatonella morbi\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eFretibacterium fastidiosum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eVeillonella_unclassified\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eGemella morbillorum\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ePrevotella nigrescens\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eStomatobaculum sp._HMT_097\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eCapnocytophaga gingivalis\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eP. endodontalis\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eTreponema socranskii\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eSelenomonas sp._HMT_126\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ePrevotella intermedia\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ePorphyromonas gingivalis\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eTreponema denticola\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eFretibacterium fastidiosum\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eTannerella sp._HMT_286\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eFretibacterium_unclassified\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eKingella_unclassified\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eBacteroidales_[G-2]_HMT_274\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003ePrevotella sp._HMT_300\u003c/em\u003e\u003c/p\u003e \u003cp\u003e\u003cem\u003eMycoplasma faucium\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eGG: generalized gingivitis, GP: generalized periodontitis, LG: localized gingivitis, LP: localized periodontitis.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eSalivary biomarkers, disease severity, and microbiome in diabetes\u003c/h2\u003e \u003cp\u003eSalivary concentrations of host response analytes by study group and periodontal disease category are depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. Here, nine host biomarkers showed major concentration differences between the NP subjects and DWP individuals, both GP and LP. MMP-8, MMP-9, TIMP-1, and MIP-1α were the only markers that significantly differed between health and the subgroups of gingivitis (DWoP) patients. Resistin concentrations were significantly increased in the DWP periodontitis subgroups, while differences in adiponectin levels between disease categories were not observed.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e As the oral microbiome both drives and responds to host responses in the oral cavity, it was of interest to explore the relationship of targeted salivary biomolecules to the microbiome components and how that relationship was affected by diabetes and periodontal characteristics. Figure\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA shows the frequency of significant correlations of the individual biomarker levels with the relative abundance of the 104 OTUs in the microbiome samples. These correlations were generally higher in the NP and DWP groups, with MMP-8, BAFF, adiponectin, and resistin showing the greatest frequency of positive correlations within the 3 subject groups. In contrast, there were only a limited number of negative correlations with any of the host molecules and the array of microbial taxa identified. Figure\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB summarizes the details of the bacterial species that dominated these correlations with the 11 salivary biomarkers. As was expected few correlations with individual bacterial species were noted in the DWoP samples. The findings depicts 27 of the OTUs with extensive correlations to the salivary analytes, with 17 in the DWP patients and 10 in the NP subjects. The identified OTUs encompassed members of the \u003cem\u003eBacteroidetes, Firmicutes (Bacillota), Fusobacterium, Spirochaetes\u003c/em\u003e, and \u003cem\u003eSynergistetes\u003c/em\u003e phyla.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eSupplemental Fig.\u0026nbsp;2A-D\u003c/b\u003e summarizes features of the correlations between individual OTUs of the 104 dominant members and levels of the individual salivary analytes. In the DWoP LG group, several significant correlations were observed, with the majority of positive correlations occurring with resistin and TIMP-1 (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). In the DWoP GG patients, IL-1\u0026szlig; and adiponectin showed parallel patterns of positive correlations (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Additionally, correlations with MMP-8 and IL-6 were all positive but with different OTUs. All significant correlations with MIP-1α were negative in this patient subset. Figure\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eC summarizes the values for the DWP LP group that showed IFNα, TIMP-1, and adiponectin positively correlated with multiple OTUs. Finally, the largest frequency of correlations was noted in the DWP GP (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD) subjects. Here IL-1β, IL-6, IFNα, BAFF, adiponectin, and resistin were the predominant biomarkers that correlated with specific OTU, and frequently the same OTU.\u003c/p\u003e \u003cp\u003e These various relationships allowed us to explore whether a targeted array of microbes and salivary biomarkers might be predictive of generalized and localized disease in T2DM patients. Odds Ratios (OR) were determined comparing levels of bacteria and host response biomolecules in periodontitis versus gingivitis subjects (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA). Here we observed OR of \u0026gt;\u0026thinsp;2.5 for most of the host biomarkers in the GP group, as well as increased ORs in the LP group. A similar number of oral bacteria exhibited ORs\u0026thinsp;\u0026gt;\u0026thinsp;2 in LP or GP when compared to the gingivitis group. However, striking was an array of the bacterial species that displayed substantially lower ORs in the GP group compared to GG group. Figure\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eB provides a similar analysis for generalized versus localized inflammation or disease. Over 60% of the targeted host analytes showed ORs\u0026thinsp;\u0026gt;\u0026thinsp;2 in LP compared with GP. Additionally, multiple bacterial species were specifically elevated in samples in the GP patients (ORs\u0026thinsp;\u0026gt;\u0026thinsp;2). Similarly, multiple species of bacteria were specifically increased in the GG versus LG patients that were frequently different from those elevated in GP (eg. \u003cem\u003eR. mucilaginosa, Prevotella\u003c/em\u003e sp. HMT305, \u003cem\u003eV. dispar\u003c/em\u003e). Also, a limited number of host response markers varied in the GG compared to the LG group (i.e., IL-6, MMP-8, adiponectin).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eEpidemiological studies confirm that diabetes is a significant risk factor for periodontitis, and periodontitis increases the risk of poor diabetes management and sequelae in affected individuals.\u003csup\u003e16,39-42\u003c/sup\u003e \u0026nbsp;Thus, this interplay between these chronic inflammatory conditions reflects the difficulty in controlling glucose levels and the initiation and progression of periodontitis.\u003csup\u003e43-49\u003c/sup\u003e The related clinical features reflect dysregulated inflammatory responses that are linked to tissue changes in\u0026nbsp;T2DM, a microbiome contribution from periodontitis, and genetic regulation of the inflammatory status of the individual. l\u003csup\u003e43,50-55\u003c/sup\u003e\u0026nbsp; Extensive reports identify the bidirectional linkage of diabetes and periodontitis, and studies that identify biological factors that contribute to the various stages of periodontal disease are not well characterized with these comorbidities. Hence, we sought to further characterize the salivary microbiome and associated host responses in T2DM patients (i.e., T2DM patients with periodontitis [DWP] or with gingivitis [DWoP]), with the goal of better understanding the panel of biological factors involved in the different extent and categories of periodontal disease compared to a NP healthy control group. \u0026nbsp;This report thus described differences in the microbiomes at the individual patient level regarding the phyla distribution, and significant differences between localized and generalized gingivitis and periodontitis. \u0026nbsp; Of the 104 dominant OTUs, approximately \u0026frac14; of these bacterial identifiers showed significant differences between health and the various disease presentation of localized and generalized periodontal diseases. \u0026nbsp;These findings are consistent with the pathogenic microbiome in periodontitis, and particularly with more generalized disease. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eConsiderable variation in inflammatory biomolecules concentrations have been observed in gingival crevicular fluid, saliva, and blood of patients with diabetes and periodontal disease.\u003csup\u003e56-60\u003c/sup\u003e\u0026nbsp; Other studies have attempted to associate these mediators with periodontitis and the transition from health to metabolic syndrome and even T2DM.\u003csup\u003e61\u003c/sup\u003e\u0026nbsp; Substantial evidence attributes pathogenic mechanisms linking periodontitis and diabetes to dysregulated inflammation.\u003csup\u003e62\u003c/sup\u003e\u0026nbsp; We have shown previously in this population differences in salivary biomarkers between normal subjects and patients with T2DM, with or without periodontitis\u0026nbsp;\u003csup\u003e29\u003c/sup\u003e that was consistent with the extant literature. \u0026nbsp;However, in spite of the large number of studies on\u0026nbsp;inflammatory mechanisms in periodontitis and diabetes, few have documented oral microbiome features that relate with specific host responses in the presence of these comorbidities. Various classic periodontal pathogens have been identified in diabetic and non-diabetic periodontitis patients, with \u003cem\u003eP. gingivalis\u0026nbsp;\u003c/em\u003eappearing in higher levels in both T2DM and T1DM patients.\u003csup\u003e63-67\u003c/sup\u003e\u0026nbsp; A growing body of evidence suggests that diabetes may alter the local periodontal pocket environment favoring certain bacterial species to emerge. \u0026nbsp;However, existing studies suggest some subtle differences in the oral microbiomes in diabetic patients, but clear clinical relevance of these differences have not been discerned. \u0026nbsp;In the present study,\u0026nbsp;significant differences were observed in various salivary analytes, especially MMP-8, MMP-9, TIMP-1, IL-1b, BAFF, and resistin primarily in the DWP versus both DWoP and NP subjects. \u0026nbsp; Exploration of the relationship between the salivary analytes and individual members of the oral microbiome showed a greater number of significant, generally positive, correlations with MMP-8, BAFF, IFNa, adiponectin, and resistin in the DWP individuals. \u0026nbsp;As the relationship between the chronic diseases of T2DM and periodontitis have emphasized a dysregulated host response, these findings support alterations in the host-microbe interactions when these diseases are coincident.\u003c/p\u003e\n\u003cp\u003eOur previous report described differences in the oral microbiomes of normal subjects, T2DM patients without periodontitis (DWoP), and T2DM patients with periodontitis (DWP) (Ebersole et al., in review). \u0026nbsp;In this analysis, we also identified several differences in the microbiomes within the DWoP and DWP groups that were stratified based upon localized or generalized clinical presentation of inflammation and destructive disease. While members of the \u003cem\u003eFirmicutes (Bacillota)\u003c/em\u003e phyla were the dominant group of bacteria in both DWP and DWoP patients, there were clear differences in the GG (DWoP) and GP (DWP) subgroups showing elevated abundance of\u0026nbsp;\u003cem\u003eBacteroidetes, Fusobacteria, Spirochaetes,\u0026nbsp;\u003c/em\u003eand \u003cem\u003eSynergistetes\u003c/em\u003e compared to the localized disease group. Thus, for the first time we demonstrated within the T2DM stratification, microbiomic variations appeared to relate to the magnitude (extent, severity) of oral inflammation and disease.\u003c/p\u003e\n\u003cp\u003eFor oral health in systemically healthy subjects or T2DM patients, there is a critical balance between tissue homeostasis and a transition into the microbial driven disease of periodontitis. \u0026nbsp;Both genetic and environmental features regulate features of an individual\u0026rsquo;s response to the burden and quality of the oral microbiome.\u003csup\u003e42,68-70\u003c/sup\u003e\u0026nbsp; As we have shown previously, there are distinctive differences in targeted host response biomolecules in saliva of these T2DM patients, as well as the characteristics of the salivary microbiome patterns coincident with these altered host response profiles.\u003csup\u003e33\u003c/sup\u003e \u0026nbsp;We now identified clear differences in both the microbiome and salivary analytes related to disease extent in T2DM patients. \u0026nbsp;Additionally, correlations between selected salivary analytes and specific members of the microbiome appeared to vary with disease extent, with the T2DM patients who had GP showing the greatest number of significant relationships. \u0026nbsp;This type of analysis does not provide any cause-and-effect understanding of the processes that occur to reach this disease susceptible milieu in the oral environment. \u0026nbsp;However, the results appear to reflect certain bacterial genera/species including\u0026nbsp;\u003cem\u003eActinomyces\u003c/em\u003e_unclassified, \u003cem\u003ePr. dentalis, F. nucleatum\u003c/em\u003e_ssp._\u003cem\u003evincentii, Leptotrichia\u003c/em\u003e sp._HMT_218, \u003cem\u003eP. endodontalis, Sneathia\u003c/em\u003e_unclassified, \u003cem\u003eT. denticola\u003c/em\u003e, and \u003cem\u003eM. faucium\u003c/em\u003e that relate more directly to specific salivary analytes, such as IL-6, adiponectin, BAFF, and resistin.\u0026nbsp;\u0026nbsp;Consideration of these host and microbial biomarkers could be useful in portending the likelihood of more severe disease and aid in improved clinical decision-making on patient specific therapy.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Limitations of this study include an overall number of subjects that were stratified based upon periodontal disease clinical characteristics of health or extent of oral disease that reduced the individual subgroup size. As such, the findings observed from these small subgroups may not be biologically generalizable to the broader T2DM or non-T2DM population. \u0026nbsp; Also,\u0026nbsp;poorly controlled diabetics were not included and may manifest distinctive host and microbiome traits.\u003csup\u003e50,71\u003c/sup\u003e\u0026nbsp; This study, as most oral microbiome reports, describe the microbial ecology based upon a discrete sample thus lacking a general assessment of the permanency or transitory nature of the salivary microbiome in individual subjects. \u0026nbsp;Finally, although saliva reflected many microbiome patterns apparent in the subgingival environment, saliva may not reflect all aspects important for understanding the biology of periodontitis. \u0026nbsp;\u003c/p\u003e"},{"header":"METHODS","content":"\u003cp\u003e\u003cstrong\u003e\u003cem\u003eStudy and Participants\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis cohort clinical study, approved by the Institutional Review Board of the University of Kentucky (UK) #17-0439-F6A in accordance with the Helsinki Declaration, was performed from June 2017 to July 2019. Inclusion criteria were: 35-80 years, a minimum of 16 teeth (excluding 3rd molars), in good systemic health (excluding the case definition), or diagnosed with T2DM based on the criteria of the American Diabetes Association.\u003csup\u003e28\u003c/sup\u003e\u0026nbsp; We have reported previously that T2DM was confirmed by the patient\u0026apos;s physician with glycated hemoglobin A1c (HbA1c), body mass index (BMI), and complete head, neck, and oral examination were recorded\u0026nbsp;\u003csup\u003e29\u003c/sup\u003e. \u0026nbsp;All subjects provided signed informed consent reviewed and accepted by the University of Kentucky Institutional Review Board.\u003c/p\u003e\n\u003cp\u003eA full-mouth periodontal examination was conducted by a single calibrated dental examiner. Periodontal health measures included bleeding on probing (BOP), pocket depth (PD) and clinical attachment loss (CAL) at six sites per tooth.\u003csup\u003e30\u003c/sup\u003e\u0026nbsp; Three cohorts were enrolled: (a) T2DM subjects with chronic periodontitis (DWP; n = 29), (b) T2DM subjects without chronic periodontitis (DWoP; n = 32) and (c) a cohort of healthier participants designated as \u0026lsquo;Not-Periodontitis\u0026rsquo; (NP) (n = 31). \u0026nbsp;Periodontitis was defined as having at least four teeth in two quadrants with a probing depth (PD) of \u0026ge;5 mm, clinical attachment loss (AL) of \u0026ge;2 mm and bleeding upon probing (BOP).\u003csup\u003e31,32\u003c/sup\u003e Participants in the NP group were defined as (a) no diabetes, (b) \u0026lt;20% proximal probing sites with a bleeding score of \u0026gt; 1 (pinpoint bleeding on gentle probing), (c) \u0026lt;5% of sites with PD of \u0026ge;4 mm (d) no PD \u003cu\u003e\u0026gt;\u003c/u\u003e5 mm, (e) \u0026lt;2% of sites with CAL of \u0026gt; 2 mm with concurrent BOP and PD of \u003cu\u003e\u0026gt;\u003c/u\u003e4 mm. The three groups were balanced for age and sex.\u003c/p\u003e\n\u003cp\u003eThe extent of gingivitis and periodontitis in each patient was evaluated and used to categorize the patients as displaying localized / generalized gingivitis (LG, GG) or localized / generalized periodontitis (LP, GP). \u0026nbsp;The DWoP group demonstrated gingivitis (i.e., BOP at \u003cu\u003e\u0026lt;\u003c/u\u003e20% of sites [LG] or \u0026gt;20% BOP sites [GG] coupled with all having less than five sites with \u003cu\u003e\u0026gt;\u003c/u\u003e4 mm periodontal pockets). \u0026nbsp;The DWP group demonstrated either LP (4-87% BOP; \u003cu\u003e\u0026lt;\u003c/u\u003e10% sites with \u003cu\u003e\u0026gt;\u003c/u\u003e4 mm pockets) or GP (21-97% BOP; \u0026gt;10% sites with \u003cu\u003e\u0026gt;\u003c/u\u003e4 mm pockets).\u003c/p\u003e\n\u003cp\u003eAs reported previously\u0026nbsp;\u003csup\u003e33\u003c/sup\u003e, medical and dental histories were obtained and subjects were excluded with alcoholism; liver, kidney or salivary gland dysfunction; inflammatory bowel disease; granulomatous diseases; immunosuppressive or cancer therapy. Additionally, acute illness (\u003cem\u003ei.e.,\u003c/em\u003e fever, sore throat, body aches and diarrhea), pregnancy or lactation, use of antibiotics within the last 6 months, need for antibiotics for dental procedures, or the presence of an oral mucosal inflammatory conditions (e.g., aphthous, lichen planus, leukoplakia, and oral cancer) and current smoker were exclusion criteria.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eBiologic samples and analysis\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUnstimulated whole saliva samples was obtained from each participant, managed, stored, and\u0026nbsp;concentrations of interleukin (IL)-1\u0026beta;, interleukin-6 (IL-6), MIP-1a\u0026nbsp;(macrophage inflammatory protein 1 alpha), BAFF (B-cell activating factor; TNFSF13b), IFNa (interferon alpha), adiponectin, and resistin were measured in duplicate using Luminex technology with\u0026nbsp;human cytokine/chemokine multiplex kits (Millipore, St. Charles, MO, USA). Salivary concentrations of matrix metalloprotease (MMP)-8, MMP-9 and TIMP-1 (tissue inhibitor of matrix matellaoproteinases-1) were determined in duplicate for each subject using human Quantikine enzyme-linked immunosorbent assay kits (R\u0026amp;D Systems, Minneapolis, MN, USA,\u0026nbsp;as we have previously described.\u003csup\u003e29,34,35\u003c/sup\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSalivary microbiome characteristics were determined in each subject\u0026rsquo;s samples using 16S rRNA sequencing as we have previously described.\u003csup\u003e36-38\u003c/sup\u003e\u0026nbsp; Sequences were assigned to their respective taxonomic classification using the Human Oral Microbiome Database (HOMD V13) (http://www.homd.org/index.php?name=seqDownload\u0026amp;file\u0026amp;type=R). \u0026nbsp;The raw data are deposited at BioProject ID PRJNA516659 through the NIH NCBI.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eStatistical Analysis\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDescriptive statistics were performed for the clinical features, Operational Taxonomic Unit (OTU) microbiome differences, and log transformed salivary analyte comparisons through ANOVA or two sample t-test. Tukey\u0026rsquo; honest significant difference method (HSD) was used for pairwise comparisons of the variables among groups of patients, if an ANOVA test was significant. \u0026nbsp;The statistical software package (SAS 9.4, Cary, NC; IBM Inc., 2020) was used for the analysis and the statistical significance level was set at 0.05. \u0026nbsp;Correlation analyses were evaluated using a Pearson correlation coefficient with a significant level of 0.05. \u0026nbsp;The data for gene expression have been uploaded into GEO accession GSE180588 (https://www.ncbi.nlm.nih.gov/gds).\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eACKNOWLEDGEMENTS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank Sabrina Braden, Dawn Dawson and Jason Stevens for their contributions in the recruitment, enrollment, data collection and laboratory analyses. \u0026nbsp;This study was supported by the Delta Dental Foundation of Michigan.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthor contributions include JLE and CSM contributed to conception, design, data acquisition and interpretation, drafted and critically revised the manuscript. SSK contributed to data acquisition, analysis, and interpretation, and critically reviewed the manuscript. DDIII contributed to obtaining and interpreting the clinical data and critically reviewed the manuscript. \u0026nbsp;XDZ provided biostatistical evaluation for data analysis, interpretation, and critical revision of the manuscript. \u0026nbsp;All authors gave their final approval and agreement to be accountable for all aspects of the work. \u0026nbsp;The authors state no conflict with any information provided in the report.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe microbiome data are deposited at BioProject ID PRJNA516659 through the NIH NCBI. \u0026nbsp;The data for gene expression have been uploaded into GEO accession GSE180588 (https://www.ncbi.nlm.nih.gov/gds).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eRohm, T. V., Meier, D. T., Olefsky, J. M. \u0026amp; Donath, M. Y. Inflammation in obesity, diabetes, and related disorders. Immunity 55, 31\u0026ndash;55, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.immuni.2021.12.013\u003c/span\u003e\u003cspan address=\"10.1016/j.immuni.2021.12.013\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGeisinger, M. L. \u003cem\u003eet al.\u003c/em\u003e Systemic Inflammatory Biomarkers and Their Association With Periodontal and Diabetes-Related Factors in the Diabetes and Periodontal Therapy Trial, A Randomized Controlled Trial. J Periodontol 87, 900\u0026ndash;913, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1902/jop.2016.150727\u003c/span\u003e\u003cspan address=\"10.1902/jop.2016.150727\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGirard, D. \u0026amp; Vandiedonck, C. How dysregulation of the immune system promotes diabetes mellitus and cardiovascular risk complications. Front Cardiovasc Med 9, 991716, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fcvm.2022.991716\u003c/span\u003e\u003cspan address=\"10.3389/fcvm.2022.991716\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVelikova, T. V., Kabakchieva, P. P., Assyov, Y. S. \u0026amp; Georgiev Tcapital A, C. Targeting Inflammatory Cytokines to Improve Type 2 Diabetes Control. \u003cem\u003eBioMed research international\u003c/em\u003e 2021, 7297419, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1155/2021/7297419\u003c/span\u003e\u003cspan address=\"10.1155/2021/7297419\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePrattichizzo, F. \u003cem\u003eet al.\u003c/em\u003e Inflammageing and metaflammation: The yin and yang of type 2 diabetes. Ageing research reviews 41, 1\u0026ndash;17, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.arr.2017.10.003\u003c/span\u003e\u003cspan address=\"10.1016/j.arr.2017.10.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGokhale, N. H. \u003cem\u003eet al.\u003c/em\u003e Resistin levels in gingival crevicular fluid of patients with chronic periodontitis and type 2 diabetes mellitus. J Periodontol 85, 610\u0026ndash;617, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1902/jop.2013.130092\u003c/span\u003e\u003cspan address=\"10.1902/jop.2013.130092\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMohan, M. \u003cem\u003eet al.\u003c/em\u003e Impact of scaling and root planing on C-reactive protein levels in gingival crevicular fluid and serum in chronic periodontitis patients with or without diabetes mellitus. J Periodontal Implant Sci 44, 158\u0026ndash;168, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5051/jpis.2014.44.4.158\u003c/span\u003e\u003cspan address=\"10.5051/jpis.2014.44.4.158\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePoston, C. J. \u003cem\u003eet al.\u003c/em\u003e Statin intake is associated with MMP-1 level in gingival crevicular fluid of patients with periodontitis. Oral Dis 22, 438\u0026ndash;444, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/odi.12474\u003c/span\u003e\u003cspan address=\"10.1111/odi.12474\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTechatanawat, S. \u003cem\u003eet al.\u003c/em\u003e Salivary and serum interleukin-17A and interleukin-18 levels in patients with type 2 diabetes mellitus with and without periodontitis. PLoS One 15, e0228921, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pone.0228921\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0228921\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHirtz, C. \u003cem\u003eet al.\u003c/em\u003e The potential impact of salivary peptides in periodontitis. Crit Rev Clin Lab Sci 58, 479\u0026ndash;492, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/10408363.2021.1907298\u003c/span\u003e\u003cspan address=\"10.1080/10408363.2021.1907298\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNibali, L. \u003cem\u003eet al.\u003c/em\u003e Patterns of subgingival microbiota in different periodontal phenotypes. J Dent 117, 103912, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jdent.2021.103912\u003c/span\u003e\u003cspan address=\"10.1016/j.jdent.2021.103912\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJohnston, W. \u003cem\u003eet al.\u003c/em\u003e Mechanical biofilm disruption causes microbial and immunological shifts in periodontitis patients. Scientific reports 11, 9796, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/s41598-021-89002-z\u003c/span\u003e\u003cspan address=\"10.1038/s41598-021-89002-z\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang, S., Yu, N. \u0026amp; Arce, R. M. Periodontal inflammation: Integrating genes and dysbiosis. Periodontol 2000 82, 129\u0026ndash;142, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12267\u003c/span\u003e\u003cspan address=\"10.1111/prd.12267\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNa, H. S. \u003cem\u003eet al.\u003c/em\u003e Identification of Potential Oral Microbial Biomarkers for the Diagnosis of Periodontitis. J Clin Med 9, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/jcm9051549\u003c/span\u003e\u003cspan address=\"10.3390/jcm9051549\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCurtis, M. A., Diaz, P. I. \u0026amp; Van Dyke, T. E. The role of the microbiota in periodontal disease. Periodontol 2000 83, 14\u0026ndash;25, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12296\u003c/span\u003e\u003cspan address=\"10.1111/prd.12296\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePolak, D., Sanui, T., Nishimura, F. \u0026amp; Shapira, L. Diabetes as a risk factor for periodontal disease-plausible mechanisms. Periodontol 2000 83, 46\u0026ndash;58, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12298\u003c/span\u003e\u003cspan address=\"10.1111/prd.12298\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSudhakara, P., Gupta, A., Bhardwaj, A. \u0026amp; Wilson, A. Oral Dysbiotic Communities and Their Implications in Systemic Diseases. Dent J (Basel) 6, 10, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/dj6020010\u003c/span\u003e\u003cspan address=\"10.3390/dj6020010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSabharwal, A. \u003cem\u003eet al.\u003c/em\u003e The salivary microbiome of diabetic and non-diabetic adults with periodontal disease. J Periodontol, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/JPER.18-0167\u003c/span\u003e\u003cspan address=\"10.1002/JPER.18-0167\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePascale, A. \u003cem\u003eet al.\u003c/em\u003e Microbiota and metabolic diseases. Endocrine 61, 357\u0026ndash;371, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s12020-018-1605-5\u003c/span\u003e\u003cspan address=\"10.1007/s12020-018-1605-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNibali, L. \u003cem\u003eet al.\u003c/em\u003e Differences in the periodontal microbiome of successfully treated and persistent aggressive periodontitis. J Clin Periodontol 47, 980\u0026ndash;990, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcpe.13330\u003c/span\u003e\u003cspan address=\"10.1111/jcpe.13330\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLu, H. \u003cem\u003eet al.\u003c/em\u003e Well-maintained patients with a history of periodontitis still harbor a more dysbiotic microbiome than health. J Periodontol 91, 1584\u0026ndash;1594, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/JPER.19-0498\u003c/span\u003e\u003cspan address=\"10.1002/JPER.19-0498\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu, G. \u003cem\u003eet al.\u003c/em\u003e Shift in the subgingival microbiome following scaling and root planing in generalized aggressive periodontitis. J Clin Periodontol 45, 440\u0026ndash;452, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcpe.12862\u003c/span\u003e\u003cspan address=\"10.1111/jcpe.12862\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSimpson, T. C. \u003cem\u003eet al.\u003c/em\u003e Treatment of periodontitis for glycaemic control in people with diabetes mellitus. \u003cem\u003eThe Cochrane database of systematic reviews\u003c/em\u003e 4, CD004714, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/14651858.CD004714.pub4\u003c/span\u003e\u003cspan address=\"10.1002/14651858.CD004714.pub4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJain, P. \u003cem\u003eet al.\u003c/em\u003e Periodontitis and Systemic Disorder-An Overview of Relation and Novel Treatment Modalities. Pharmaceutics 13, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/pharmaceutics13081175\u003c/span\u003e\u003cspan address=\"10.3390/pharmaceutics13081175\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKapellas, K. \u003cem\u003eet al.\u003c/em\u003e Periodontal therapy and glycaemic control among individuals with type 2 diabetes: reflections from the PerioCardio study. International journal of dental hygiene, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/idh.12234\u003c/span\u003e\u003cspan address=\"10.1111/idh.12234\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMichalowicz, B. S. \u003cem\u003eet al.\u003c/em\u003e Factors associated with the clinical response to nonsurgical periodontal therapy in people with type 2 diabetes mellitus. J Am Dent Assoc 145, 1227\u0026ndash;1239, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.14219/jada.2014.92\u003c/span\u003e\u003cspan address=\"10.14219/jada.2014.92\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGaikwad, S. P., Gurav, A. N., Shete, A. R. \u0026amp; Desarda, H. M. Effect of scaling and root planing combined with systemic doxycycline therapy on glycemic control in diabetes mellitus subjects with chronic generalized periodontitis: a clinical study. J Periodontal Implant Sci 43, 79\u0026ndash;86, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5051/jpis.2013.43.2.79\u003c/span\u003e\u003cspan address=\"10.5051/jpis.2013.43.2.79\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAmerican, D. A. \u003cem\u003eDiagnosis\u003c/em\u003e, \u0026lt; \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.diabetes.org/a1c/diagnosis\u0026gt;\u003c/span\u003e\u003cspan address=\"https://www.diabetes.org/a1c/diagnosis%3E\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiller, C. S., Ding, X., Dawson, D. R., 3rd \u0026amp; Ebersole, J. L. Salivary biomarkers for discriminating periodontitis in the presence of diabetes. J Clin Periodontol 48, 216\u0026ndash;225, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcpe.13393\u003c/span\u003e\u003cspan address=\"10.1111/jcpe.13393\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDawson, D. R., 3rd \u003cem\u003eet al.\u003c/em\u003e Salivary levels of Epstein-Barr virus DNA correlate with subgingival levels, not severity of periodontitis. \u003cem\u003eOral Dis\u003c/em\u003e 15, 554\u0026ndash;559, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1601-0825.2009.01585.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1601-0825.2009.01585.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArmitage, G. C. Development of a classification system for periodontal diseases and conditions. Annals of periodontology / the American Academy of Periodontology 4, 1\u0026ndash;6, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1902/annals.1999.4.1.1\u003c/span\u003e\u003cspan address=\"10.1902/annals.1999.4.1.1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (1999).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArmitage, G. C., Research, S. \u0026amp; Therapy Committee of the American Academy of, P. Diagnosis of periodontal diseases. J Periodontol 74, 1237\u0026ndash;1247, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1902/jop.2003.74.8.1237\u003c/span\u003e\u003cspan address=\"10.1902/jop.2003.74.8.1237\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2003).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiller, C. S., Ding, X., Nagarajan, R., Dawson, D. R., 3rd \u0026amp; Ebersole, J. L. Biomarker panel discriminates diabetics with and without periodontitis pre- and post-therapy. J Periodontal Res 58, 493\u0026ndash;502, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jre.13127\u003c/span\u003e\u003cspan address=\"10.1111/jre.13127\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNagarajan, R., Miller, C. S., Dawson, D., 3rd, Al-Sabbagh, M. \u0026amp; Ebersole, J. L. Patient-Specific Variations in Biomarkers across Gingivitis and Periodontitis. PLoS One 10, e0136792, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pone.0136792\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0136792\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEbersole, J. L., Nagarajan, R., Akers, D. \u0026amp; Miller, C. S. Targeted salivary biomarkers for discrimination of periodontal health and disease(s). Frontiers in cellular and infection microbiology 5, 62, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fcimb.2015.00062\u003c/span\u003e\u003cspan address=\"10.3389/fcimb.2015.00062\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKirakodu, S., Chen, J., Gonzalez Martinez, J., Gonzalez, O. A. \u0026amp; Ebersole, J. Microbiome Profiles of Ligature-Induced Periodontitis in Nonhuman Primates across the Life Span. Infect Immun 87, e00067-00019, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1128/IAI.00067-19\u003c/span\u003e\u003cspan address=\"10.1128/IAI.00067-19\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKirakodu, S. S., Govindaswami, M., Novak, M. J., Ebersole, J. L. \u0026amp; Novak, K. F. Optimizing qPCR for the Quantification of Periodontal Pathogens in a Complex Plaque Biofilm. Open Dent J 2, 49\u0026ndash;55, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.2174/1874210600802010049\u003c/span\u003e\u003cspan address=\"10.2174/1874210600802010049\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEbersole, J. L., Kirakodu, S. S., Dawson III, D., Nagarajan, R. \u0026amp; Miller, C. S. Microbiome characteristics of diabetics with periodontitis before and after treatment. Molecular Oral MIcrobiology in review (2024).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePussinen, P. J. \u003cem\u003eet al.\u003c/em\u003e Periodontitis and cardiometabolic disorders: The role of lipopolysaccharide and endotoxemia. Periodontol 2000 89, 19\u0026ndash;40, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12433\u003c/span\u003e\u003cspan address=\"10.1111/prd.12433\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBhuyan, R. \u003cem\u003eet al.\u003c/em\u003e Periodontitis and Its Inflammatory Changes Linked to Various Systemic Diseases: A Review of Its Underlying Mechanisms. Biomedicines 10, 2659, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/biomedicines10102659\u003c/span\u003e\u003cspan address=\"10.3390/biomedicines10102659\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBains, V. K., Mahendra, J., Mahendra, L., Mittal, M. \u0026amp; Valli, G. Markers, Pathways, and Current Evidence for Periodontitis-associated Insulin Resistance: A Narrative Review. Journal of International Society of Preventive \u0026amp; Community Dentistry 12, 475\u0026ndash;487, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4103/jispcd.JISPCD_92_22\u003c/span\u003e\u003cspan address=\"10.4103/jispcd.JISPCD_92_22\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKapila, Y. L. Oral health's inextricable connection to systemic health: Special populations bring to bear multimodal relationships and factors connecting periodontal disease to systemic diseases and conditions. Periodontol 2000 87, 11\u0026ndash;16, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12398\u003c/span\u003e\u003cspan address=\"10.1111/prd.12398\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNascimento, G. G., Leite, F. R. M., Vestergaard, P., Scheutz, F. \u0026amp; Lopez, R. Does diabetes increase the risk of periodontitis? A systematic review and meta-regression analysis of longitudinal prospective studies. Acta Diabetol 55, 653\u0026ndash;667, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s00592-018-1120-4\u003c/span\u003e\u003cspan address=\"10.1007/s00592-018-1120-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGenco, R. J. \u0026amp; Borgnakke, W. S. Risk factors for periodontal disease. Periodontol 2000 62, 59\u0026ndash;94, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1600-0757.2012.00457.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1600-0757.2012.00457.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLalla, E. \u003cem\u003eet al.\u003c/em\u003e Periodontal Changes in Children and Adolescents With Diabetes: A case-control study. Diabetes Care 29, 295\u0026ndash;299 (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDemmer, R. T., Jacobs, D. R., Jr. \u0026amp; Desvarieux, M. Periodontal disease and incident type 2 diabetes: results from the First National Health and Nutrition Examination Survey and its epidemiologic follow-up study. Diabetes Care 31, 1373\u0026ndash;1379, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.2337/dc08-0026\u003c/span\u003e\u003cspan address=\"10.2337/dc08-0026\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBorgnakke, W. S., Ylostalo, P. V., Taylor, G. W. \u0026amp; Genco, R. J. Effect of periodontal disease on diabetes: systematic review of epidemiologic observational evidence. J Clin Periodontol 40 Suppl 14, S135-152, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcpe.12080\u003c/span\u003e\u003cspan address=\"10.1111/jcpe.12080\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlbert, D. A. \u003cem\u003eet al.\u003c/em\u003e Diabetes and oral disease: implications for health professionals. Ann N Y Acad Sci 1255, 1\u0026ndash;15, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1749-6632.2011.06460.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1749-6632.2011.06460.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChapple, I. L., Genco, R. \u0026amp; working group 2 of the joint, E. F. P. A. A. P. w. Diabetes and periodontal diseases: consensus report of the Joint EFP/AAP Workshop on Periodontitis and Systemic Diseases. J Periodontol 84, S106-112, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1902/jop.2013.1340011\u003c/span\u003e\u003cspan address=\"10.1902/jop.2013.1340011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSanz, M. \u003cem\u003eet al.\u003c/em\u003e Scientific evidence on the links between periodontal diseases and diabetes: Consensus report and guidelines of the joint workshop on periodontal diseases and diabetes by the International Diabetes Federation and the European Federation of Periodontology. J Clin Periodontol 45, 138\u0026ndash;149, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcpe.12808\u003c/span\u003e\u003cspan address=\"10.1111/jcpe.12808\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNguyen, A. T. M. \u003cem\u003eet al.\u003c/em\u003e The association of periodontal disease with the complications of diabetes mellitus. A systematic review. Diabetes Res Clin Pract 165, 108244, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.diabres.2020.108244\u003c/span\u003e\u003cspan address=\"10.1016/j.diabres.2020.108244\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGraziani, F., Gennai, S., Solini, A. \u0026amp; Petrini, M. A systematic review and meta-analysis of epidemiologic observational evidence on the effect of periodontitis on diabetes An update of the EFP-AAP review. J Clin Periodontol 45, 167\u0026ndash;187, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcpe.12837\u003c/span\u003e\u003cspan address=\"10.1111/jcpe.12837\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZiukaite, L., Slot, D. E., Cobb, C. M., Coucke, W. \u0026amp; Van der Weijden, G. A. Prevalence of diabetes among patients diagnosed with periodontitis: A retrospective cross-sectional study. International journal of dental hygiene 16, 305\u0026ndash;311, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/idh.12280\u003c/span\u003e\u003cspan address=\"10.1111/idh.12280\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChapple, I. L. \u003cem\u003eet al.\u003c/em\u003e Interaction of lifestyle, behaviour or systemic diseases with dental caries and periodontal diseases: consensus report of group 2 of the joint EFP/ORCA workshop on the boundaries between caries and periodontal diseases. J Clin Periodontol 44 Suppl 18, S39-S51, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcpe.12685\u003c/span\u003e\u003cspan address=\"10.1111/jcpe.12685\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePolak, D. \u0026amp; Shapira, L. An update on the evidence for pathogenic mechanisms that may link periodontitis and diabetes. J Clin Periodontol 45, 150\u0026ndash;166, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcpe.12803\u003c/span\u003e\u003cspan address=\"10.1111/jcpe.12803\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKardesler, L., Buduneli, N., Cetinkalp, S. \u0026amp; Kinane, D. F. Adipokines and inflammatory mediators after initial periodontal treatment in patients with type 2 diabetes and chronic periodontitis. J Periodontol 81, 24\u0026ndash;33, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1902/jop.2009.090267\u003c/span\u003e\u003cspan address=\"10.1902/jop.2009.090267\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKardesler, L., Buduneli, N., Cetinkalp, S., Lappin, D. \u0026amp; Kinane, D. F. Gingival crevicular fluid IL-6, tPA, PAI-2, albumin levels following initial periodontal treatment in chronic periodontitis patients with or without type 2 diabetes. Inflamm Res 60, 143\u0026ndash;151, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s00011-010-0248-7\u003c/span\u003e\u003cspan address=\"10.1007/s00011-010-0248-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKardesler, L. \u003cem\u003eet al.\u003c/em\u003e Crevicular fluid matrix metalloproteinase-8, -13, and TIMP-1 levels in type 2 diabetics. Oral Dis 16, 476\u0026ndash;481, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1601-0825.2010.01659.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1601-0825.2010.01659.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNavarro-Sanchez, A. B., Faria-Almeida, R. \u0026amp; Bascones-Martinez, A. Effect of non-surgical periodontal therapy on clinical and immunological response and glycaemic control in type 2 diabetic patients with moderate periodontitis. J Clin Periodontol 34, 835\u0026ndash;843, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1600-051X.2007.01127.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1600-051X.2007.01127.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2007).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSantos, V. R. \u003cem\u003eet al.\u003c/em\u003e Cytokine levels in sites of chronic periodontitis of poorly controlled and well-controlled type 2 diabetic subjects. J Clin Periodontol 37, 1049\u0026ndash;1058, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1600-051X.2010.01624.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1600-051X.2010.01624.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGenco, R. J., Graziani, F. \u0026amp; Hasturk, H. Effects of periodontal disease on glycemic control, complications, and incidence of diabetes mellitus. Periodontol 2000 83, 59\u0026ndash;65, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12271\u003c/span\u003e\u003cspan address=\"10.1111/prd.12271\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTaylor, J. J., Preshaw, P. M. \u0026amp; Lalla, E. A review of the evidence for pathogenic mechanisms that may link periodontitis and diabetes. J Periodontol 84, S113\u0026ndash;134, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1902/jop.2013.134005\u003c/span\u003e\u003cspan address=\"10.1902/jop.2013.134005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu, L. S., Gkranias, N., Farias, B., Spratt, D. \u0026amp; Donos, N. Differences in the subgingival microbial population of chronic periodontitis in subjects with and without type 2 diabetes mellitus-a systematic review. Clin Oral Investig 22, 2743\u0026ndash;2762, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s00784-018-2660-2\u003c/span\u003e\u003cspan address=\"10.1007/s00784-018-2660-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLira-Junior, R., Akerman, S., Klinge, B., Bostrom, E. A. \u0026amp; Gustafsson, A. Salivary microbial profiles in relation to age, periodontal, and systemic diseases. PLoS One 13, e0189374, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pone.0189374\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0189374\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eField, C. A., Gidley, M. D., Preshaw, P. M. \u0026amp; Jakubovics, N. Investigation and quantification of key periodontal pathogens in patients with type 2 diabetes. J Periodontal Res 47, 470\u0026ndash;478, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1600-0765.2011.01455.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1600-0765.2011.01455.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMakiura, N. \u003cem\u003eet al.\u003c/em\u003e Relationship of Porphyromonas gingivalis with glycemic level in patients with type 2 diabetes following periodontal treatment. Oral Microbiol Immunol 23, 348\u0026ndash;351, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1399-302X.2007.00426.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1399-302X.2007.00426.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEbersole, J. L., Holt, S. C., Hansard, R. \u0026amp; Novak, M. J. Microbiologic and immunologic characteristics of periodontal disease in Hispanic americans with type 2 diabetes. J Periodontol 79, 637\u0026ndash;646, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1902/jop.2008.070455\u003c/span\u003e\u003cspan address=\"10.1902/jop.2008.070455\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLoos, B. G. \u0026amp; Van Dyke, T. E. The role of inflammation and genetics in periodontal disease. Periodontol 2000 83, 26\u0026ndash;39, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12297\u003c/span\u003e\u003cspan address=\"10.1111/prd.12297\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDommisch, H., Kuzmanova, D., Jonsson, D., Grant, M. \u0026amp; Chapple, I. Effect of micronutrient malnutrition on periodontal disease and periodontal therapy. Periodontol 2000 78, 129\u0026ndash;153, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12233\u003c/span\u003e\u003cspan address=\"10.1111/prd.12233\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eReynolds, M. A. Modifiable risk factors in periodontitis: at the intersection of aging and disease. Periodontol 2000 64, 7\u0026ndash;19, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/prd.12047\u003c/span\u003e\u003cspan address=\"10.1111/prd.12047\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMiranda, T. S. \u003cem\u003eet al.\u003c/em\u003e Influence of glycemic control on the levels of subgingival periodontal pathogens in patients with generalized chronic periodontitis and type 2 diabetes. Journal of applied oral science: revista FOB 25, 82\u0026ndash;89, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1590/1678-77572016-0302\u003c/span\u003e\u003cspan address=\"10.1590/1678-77572016-0302\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"diabetes, microbiome, inflammation, periodontitis, saliva","lastPublishedDoi":"10.21203/rs.3.rs-4391317/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4391317/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eType 2 diabetes mellitus (T2DM) is associated with cellular abnormalities, tissue and organ dysfunctions, and periodontitis. This investigation examined the relationship between the oral microbiome and salivary biomarkers in T2DM patients with or without periodontitis. This cohort (35\u0026ndash;80 years) included systemically healthy non-periodontitis (NP; n\u0026thinsp;=\u0026thinsp;31), T2DM without periodontitis (DWoP; n\u0026thinsp;=\u0026thinsp;32) and T2DM with periodontitis (DWP; n\u0026thinsp;=\u0026thinsp;29). The oral microbiome [Operational Taxonomic Units (OTUs)] (16s rRNA sequencing) and targeted host salivary biomarkers (immunoassays) were assessed. We identified 47 OTUs that were significantly different in abundance between NP samples and any disease subset or between disease subgroups. The most unique microbiome patterns were observed in the DWP group. Differences in genera/species abundance were also observed when T2DM patients were stratified by extent of periodontal inflammation and disease (i.e., generalized versus localized gingivitis/periodontitis). Salivary biomarkers showed significant elevations in MMP-8, MMP-9, resistin, IL-1β, IL-6, IFNα, and BAFF (THFSR13b) comparing generalized to localized periodontitis. Salivary analytes showed significant positive correlations with specific microbiome members, predominantly in DWP patients. Odds ratio analyses reinforced that a panel of biologic markers (IL-6, MMP-8) and bacteria (e.g., \u003cem\u003eBacteroidetes, Fusobacteria, Spirochaetes\u003c/em\u003e) discriminated the severity and extent of periodontal disease in this diabetic population.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e","manuscriptTitle":"Salivary Features of Periodontitis and Gingivitis in Type 2 Diabetes Mellitus","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-14 18:39:12","doi":"10.21203/rs.3.rs-4391317/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-07-15T16:25:09+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-07-14T19:54:57+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"89556361353753141459695577028337487057","date":"2024-07-14T17:51:25+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-07-14T07:11:10+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"102981776773898459098876889884522258090","date":"2024-07-03T09:47:31+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-06-05T23:37:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"5397488384503131848159056166013565035","date":"2024-05-31T12:10:19+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"32346161046186761575780900844489020550","date":"2024-05-31T08:29:31+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-05-31T08:27:34+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-05-31T08:06:42+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-05-31T08:01:40+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-05-31T07:59:35+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-05-08T20:17:25+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9f19c7dd-19d1-4ada-b6fd-4ff9edd44ea4","owner":[],"postedDate":"June 14th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":33046852,"name":"Biological sciences/Immunology"},{"id":33046853,"name":"Biological sciences/Microbiology"},{"id":33046854,"name":"Health sciences/Biomarkers"},{"id":33046855,"name":"Health sciences/Diseases"}],"tags":[],"updatedAt":"2024-12-30T16:05:23+00:00","versionOfRecord":{"articleIdentity":"rs-4391317","link":"https://doi.org/10.1038/s41598-024-77434-2","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2024-12-28 15:57:41","publishedOnDateReadable":"December 28th, 2024"},"versionCreatedAt":"2024-06-14 18:39:12","video":"","vorDoi":"10.1038/s41598-024-77434-2","vorDoiUrl":"https://doi.org/10.1038/s41598-024-77434-2","workflowStages":[]},"version":"v1","identity":"rs-4391317","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4391317","identity":"rs-4391317","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}